BLASTing Flu Viruses

swimmy · Post by **swimmy** » Wed Aug 19, 2009 3:30 pm

I wanted to try the BLASTing Flu Viruses project. However, the BLAST at ISD doesn't work. Would the BLAST at NCBI be the same? If not, please tell me what is different.

The vaccines are on the ISD page link given in the project, right?

If those are the vaccines, then where would the raw sequences for the actual flu be? How could I find them?

Should I compare the Hemagluttinin protein of the vaccines and viruses or the Neuraminidase protein?

Are the vaccines ones that are currently being used?

Could I compare how similar all the vaccines are to swine flu?

I also don't really get the results that appear after trying the experimental procedure with NCBI's BLAST.

Should I just enter the DNA sequence of the vaccine, or Align two or more sequences? If I do just the DNA sequence of the vaccine, then it says "No significant similarities found" and there are no results.

NOTE: I found the Influenza Research Database, which has sequences as well as BLAST.
Here is the site:
http://www.biohealthbase.org/GSearch/bl ... =Influenza

Is this the same as ISD? There was a link that led here from ISD home page.

deleted-2574 · Post by **deleted-2574** » Fri Aug 21, 2009 9:44 am

Hi swimmy,

Wow! I'm impressed by the depth of knowledge in your post, particularly in Grades 6-8. I don't profess to know answers to your questions. Here's a method that may help getting answers. If you type "blast" into the "Search" box at the upper right of the sciencebuddies main page, you get 57 hits before yours, in the range of 8/2/2005 to 1/4/2009.

Some of these, and the pages they reference, may address the topics in your post. Do they help?

By the way, of your terms, ISD, NCBI, Hemagluttinin and Neuraminidase, only NCBI has references on sciencebuddies, 63 (more than blast!) hits besides yours. Is it a case where there's smoke, there's fire? So, ISD, Hemagluttinin and Neuraminidase may need more research.

Does this help?

swimmy · Post by **swimmy** » Fri Aug 21, 2009 1:08 pm

Some of the pages do help, but sometimes some of the people posted in many different forums, or they have different information about the things I am wondering about.

Here are questions I still need answers to:

The page with the sequences of the vaccines does not work. http://www.flu.lanl.gov/vaccine
Could I use sequences of vaccines from http://www.biohealthbase.org/GSearch/va ... =influenza instead?

Where would the raw sequences for the actual flu be? How could I find them? Are they on the NCBI page see http://www.ncbi.nlm.nih.gov/genomes/FLU/SwineFlu.html

Should I compare the Hemagluttinin protein of the vaccines and viruses or the Neuraminidase protein?

Are the vaccines (http://www.flu.lanl.gov/vaccine or http://www.biohealthbase.org/GSearch/va ... =influenza (if it can be used) ones that are currently being used?

Could I compare how similar all the vaccines are to swine flu? To find out if they are effective for the swine flu outbreak?

I also don't really get the results that appear after trying the experimental procedure with NCBI's BLAST. What percentage should be matched for the vaccine to be effective?

Should I just enter the DNA sequence of the vaccine, or Align two or more sequences? If I do just the DNA sequence of the vaccine, then it says "No significant similarities found" and there are no results.

I also don't get the "database" comparing to flu sequences. Which one should I use if any?

Also, what vaccine should I choose?

I know these are still most of the questions I asked before, but after searching "BLAST" on the search, then I still couldn't find the information to answer these questions or there was ambiguity in the answers.

MelissaB · Post by **MelissaB** » Thu Aug 27, 2009 2:49 am

Hi,

First of all, yes, you can use vaccine sequences from the site you found.

I find the CDC website (http://www.cdc.gov/flu/weekly/fluactivity.htm) to be most helpful for figuring out which flu strains went around. For example, if you look at the summary of the 2007-2008 season, it will tell you what sequence most strains were 'antigenically similar to'. You can then use the name of this strain to look up the sequence in BLAST. For example, in 2007-2008, 66% of type A infections were similar to A/Solomon Islands/3/2006.

It is up to you whether you want to compare the H protein or the N protein.

As far as I know, the WHO recommendations are indeed the vaccines that were used (at least in North America). For earlier years, the CDC pages I pointed you to above list the sequences in the vaccines.

On this site: http://www.ncbi.nlm.nih.gov/genomes/FLU/FLU.html you can find 'sequences from the pandemic (2009) viruses', which should give you some swine flu sequences to work with. So yes, you could see whether or not the vaccine sequences are close to the swine flu sequences.

As for effectiveness, you can usually get a good estimate by looking at the proportion of samples that are antigenically similar to the proteins in the vaccine. But generally, the closer the match, the more likely the vaccine will work.

It is up to you which vaccine you use! You might consider comparing the similarities between this year's normal flu vaccine, the swine flu vaccine, and the swine flu--it sounds as if you're interested in whether or not the 'normal' vaccine will work, right?

Definitely align two different sequences with BLAST--the vaccine, and the flu sample you're interested in.

Take a good look through the CDC website and the information it has, then post back if you have more questions.

swimmy · Post by **swimmy** » Sat Aug 29, 2009 6:19 pm

Thanks, this really helps.
swimmy

amyC · Post by **amyC** » Tue Sep 01, 2009 4:33 pm

Hi Swimmy - I have been in contact with the team of scientists here at Science Buddies and the author of the project you have been working on. Based on the questions you've raised, we are editing the project some to make parts of it clearer. I have some answers to pass along to you from the project author. I hope these help. I'm copying your question in here along with his answer:

1. The page with the sequences of the vaccines does not work. http://www.flu.lanl.gov/vaccine - OUT OF COMMISSION
Could I use sequences of vaccines from http://www.biohealthbase.org/GSearch/va ... =influenza instead?

Answer: Yes. But it might be easier to use the NBCI site: http://www.ncbi.nlm.nih.gov/Genbank/

2. Where would the raw sequences for the actual flu be? How could I find them? Are they on the NCBI page?

Answer: See http://www.ncbi.nlm.nih.gov/genomes/FLU/SwineFlu.html

3. Should I compare the Hemagluttinin protein of the vaccines and viruses or the Neuraminidase protein?

Answer: Use the hemagluttinin (HA) protein sequence. This is the protein that is assayed by the hemaglutination assay that is used to test how well the vaccine is working. And protein sequence is more informative that dna since dna accumulates silent mutations that complicate interpretation of the blast results.

4. Could I compare how similar all the vaccines are to swine flu? To find out if they are effective for the swine flu outbreak?

Answer: Yes. Go to the NCBI Flu page for the sequence: See http://www.ncbi.nlm.nih.gov/genomes/FLU/SwineFlu.html

5. I also don't really get the results that appear after trying the experimental procedure with NCBI's BLAST. What percentage should be matched for the vaccine to be effective?

Answer: It is hard to say up front what percentage should be matched. I suggest that you formulate a new question that the BLAST output is able to address. I have not tried any of the projects below, so they may not be good ways to proceed, but they will give you an idea of the kind of project I have in mind. For example, you could ask

A) What part of the protein is most subject to change? (Might try to form a hypothesis as to why certain parts of the protein are more subject to change). Might want to focus on one variation to make the project manageable. For example, this part of the HA protein has a change from a D to an N.

Query 121 YASLRSLVASSGTLEFNDESFNWTGVTQNGT
YASLRSLVASSGTLEFN+ESFNWTGVTQNGT
Sbjct 121 YASLRSLVASSGTLEFNNESFNWTGVTQNGT

B) How are various strains distributed geographically?

C) Can you identify when (what year) a certain variant appeared and how it spread?

6. Should I just enter the DNA sequence of the vaccine, or Align two or more sequences? If I do just the DNA sequence of the vaccine, then it says "No significant similarities found" and there are no results.

Answer: Use protein sequence.

7. I also don't get the "database" comparing to flu sequences. Which one should I use if any?

Answer: Use the protein database

8. Also, what vaccine should I choose? I would pick a year for which there is good molecular data, and not too recent. Say 2004.

Answer: It is up to you!

I hope these help, Swimmy. If you have additional questions, please post here, and I'll follow up with the scientist. I'll also post here and let everyone know when the project is officially updated.

Amy Cowen
Science Buddies

swimmy · Post by **swimmy** » Thu Sep 03, 2009 6:39 pm

Thanks, Amy.
Ok, I understand where to find the raw sequences of the actual flu virus. I checked the site (http://www.ncbi.nlm.nih.gov/Genbank/) but I don't really get how to get the flu vaccine sequence still.
I searched "Are normal season flu vaccines effective for swine flu?" on google. There are varied results, so I want to go with one of the ideas that you mentioned on #5. I want to do #5 (A), but I don't really get how to go about doing that.

By protein database, do you mean Protein Data Bank (pdb)? This is the only protein database I could find in blastn.

amyC · Post by **amyC** » Fri Sep 04, 2009 9:01 am

Hi Swimmy - I have some good news! The Project Idea has been updated with new directions that I hope will help clear up some of the trouble you ran into. The project is here:

https://www.sciencebuddies.org/science- ... p003.shtml

After looking it over, if you have questions, let us know.

Amy
Science Buddies

swimmy · Post by **swimmy** » Fri Sep 04, 2009 2:13 pm

Thanks again, Amy.

CDC has antigenically characterized 2,112 seasonal human influenza viruses [1,189 influenza A (H1), 227 influenza A (H3) and 696 influenza B viruses] collected by U.S. laboratories since October 1, 2008, and 493 2009 influenza A (H1N1) viruses.

All 1,189 seasonal influenza A (H1) viruses are related to the influenza A (H1N1) component of the 2008-09 influenza vaccine (A/Brisbane/59/2007). Two hundred sixteen (95%) of 227 influenza A (H3N2) viruses tested are related to the A (H3N2) vaccine component (A/Brisbane/10/2007) and 11 viruses (5%) tested showed reduced titers with antisera produced against A/Brisbane/10/2007.

"All 493 2009 influenza A (H1N1) viruses are related to the A/California/07/2009 (H1N1)pdm reference virus selected by WHO as a potential candidate for 2009 influenza A (H1N1) vaccine."

~CDC

This is about the H1N1 virus. It says the virus is related to a part of the vaccine for seasonly flu.
Should I use those strains mentioned as the vaccines?
Or should I use the candidate strain for the vaccine?

I tried the method in the revised edition of BLASTIng Flu Viruses. I am successful with the procedure but then it says "Putative conserved domains have been detected, click on the image below for detailed results." What does this mean?

Also, the results are kind of confusing, because I don't see the alignments...
also, there are no lowercase letters. I'm not really sure if what I am seeing is correct, if it is correct, I don't know how to analyze the results.

Here is the blast search:http://blast.ncbi.nlm.nih.gov/Blast.cgi ... _box_blast

here is the HA protein I used http://www.ncbi.nlm.nih.gov/protein/ACQ ... e_RVDocSum

Why are the letters not just A, G, C and T?
Is it the amino acids?

What do the plus signs (+) and the spaces mean?

Also how would I be able to see the rate at which antigenic shift occurs? I might use this as an idea...
Would I be able to see the effect of environment on the rate of mutation?

MelissaB · Post by **MelissaB** » Sat Sep 05, 2009 2:11 am

Swimmy,

I'll let Amy answer most of your questions, but 'putative conserved domains' just means 'places where the virus doesn't mutate'. These are generally places that have to be conserved in order for the protein to function properly--for example, the parts that anchor in into the virus' membrane, or parts relating to the protein's function.

swimmy · Post by **swimmy** » Sun Sep 06, 2009 12:19 pm

In addition to all those questions above,
I have a few questions about writing the report for this project.
If I had the title "Which regions of the influenza virus protein are most subject to change?", what could I write for my hypothesis?
Do I just specify the kind of protein? (That doesn't make much sense to me, because it is just the hemagluttinin protein that I am using)
Would I need to cite where I got the protein sequence?
and also, would i need to cite the websites in the procedure?

I need help really quickly because I need to finish a draft by tuesday, but I'll be busy tomorrow.

MelissaB · Post by **MelissaB** » Mon Sep 07, 2009 12:29 am

Hi,

Yes, you should definitely cite the websites you used during your search and tell where you got the protein sequence.

In order to form a hypothesis, you would need to do research on what the various parts of the protein do (are they structural, do they anchor the protein to the membrane, do they interact with other cells, etc.). Then you could guess whether or not they will be subject to change.

swimmy · Post by **swimmy** » Fri Sep 11, 2009 4:49 pm

ok, so i have turned in the draft with conclusion saying i didn't know if the results were correct.
so, i still need help on the question from september 4 (last week)
and i would appreciate help for the other question too

swimmy · Post by **swimmy** » Sun Sep 13, 2009 11:26 am

I have done research on different types of proteins in influenza
here are the proteins:
hemagglutinin
neuraminidase
nucleocapsid
nuclear export protein
nonstructural protein 1
Polymerase PA
polymerase PB1
polymerase PB2
matrix protein 1
matrix protein 2

these are all the proteins I could find in GenBank
so should i do blast searches for all of them and then compare how many lowercase letters (i still need help on this)?
if not, what should i do?
and also how should i figure out what region of protein? is it one of the types mentioned above or an actual region?
how would I describe it?

how do i site the sequences?
also, can you please help me understand the paper http://www.biology-direct.com/content/p ... 0-4-18.pdf

swimmy · Post by **swimmy** » Mon Sep 14, 2009 4:56 pm

I want to submit this to the school science fair...
I need help identifying the independent/manipulated, dependent/responding, and controlled/constant variables.
What should I use for a graph?

These things are on the application.

Please help on questions posted september 4th and later

amyC · Post by **amyC** » Tue Sep 15, 2009 8:17 am

Hi Swimmy - I have asked the author of the project to take a look at your questions from September 4 as you worked through the revised project. As soon as I hear back, I'll post his response here.

In the meantime, you might want to look back at the Science Buddies' information on "variables" to see if you can identify the variables in the project:
https://www.sciencebuddies.org/science- ... bles.shtml

Amy
Science Buddies

amyC · Post by **amyC** » Tue Sep 15, 2009 1:20 pm

Hi Swimmy -

I've got some answers for you.

You asked:

"This is about the H1N1 virus. It says the virus is related to a part of the vaccine for seasonly flu.
Should I use those strains mentioned as the vaccines? "

Answer: Yes, I would use the A/Brisbane/59/2007 sequence since that is a component of the 2008-9 vaccine. The protein sequence can be found here: http://www.ncbi.nlm.nih.gov/protein/ACA ... e_RVDocSum.

Click on "BLAST Sequence" on this page. A new page appears.

Click on "BLAST" on the next page to BLAST the protein against a protein database. The default database is the non-redundant database of proteins at NCBI, which includes almost every protein sequence that has been published. The default number of alignments is 100. You can click on "Algorithm parameters" to increase the number of alignments. You should spend some time looking up more about the BLAST tool using the links provided on the BLAST page, for example http://blast.ncbi.nlm.nih.gov/Blast.cgi ... =BlastDocs.

To limit the database to sequences that were found in a particular year, for example 2009, type "2009 [WORD]" in the box that says Entrez query on the BLAST page. This might be useful to look at changes over time.

The output includes identification of a "Putative Conserved Domain". The "Putative Conserved Domain" output tells you what protein family your input sequence belongs in. This is found by comparing your input sequence to a database of conserved sequences for various kinds of proteins. This is information that is included in addition to the BLAST output. BLAST just identifies similar sequences and aligns them. The input belongs in the hemagglutinin family, which makes sense since you are using the HA protein (influenza hemagglutinin) as an input. If you click on the image of the hemagglutinin you can learn more about this family of proteins. If you want to learn more about how this tool works, see this paper: http://www.pubmedcentral.nih.gov/articl ... tid=308855.

Scroll down to see the BLAST output. There are 20 amino acids in proteins. The BLAST output uses the standard 1 letter code to identify amino acids. You can learn about these here: http://www.biology.arizona.edu/biochemi ... aa/aa.html. You can find similar info on other sites on the web or a good biology textbook.

You asked:

I tried the method in the revised edition of BLASTIng Flu Viruses. I am successful with the procedure but then it says "Putative conserved domains have been detected, click on the image below for detailed results." What does this mean?

Answer: See explanation above.

Also, the results are kind of confusing, because I don't see the alignments...

Answer: See explanation above regarding how to get BLAST alignments.

You said:

also, there are no lowercase letters. I'm not really sure if what I am seeing is correct, if it is correct, I don't know how to analyze the results.

Answer: Lower case letters are used in BLAST outputs to show where sequence with low complexity, such as a sequence like AAAAAAAAAAAAA, were filtered out. These low-complexity sequences are not very informative and confuse the BLAST output interpretation. There are none in the HA sequence

You asked:

What do the plus signs (+) and the spaces mean?

Answer: Plus signs mean that the amino acid change is considered "conservative". In other words, it is a change that is often found in other proteins and probably won't disrupt the structure of the protein.

You asked about your hypothesis:

If I had the title "Which regions of the influenza virus protein are most subject to change?", what could I write for my hypothesis?

This is up to you. Might be something like "not all regions will change equally because …."

Do I just specify the kind of protein?

Yes

Would I need to cite where I got the protein sequence?

Yes

would i need to cite the websites in the procedure?

Yes

He also notes that standard controlled, dependent, and independent variables might not apply in the traditional way for the type of project you are doing.

I hope this helps!

Amy
Science Buddies

swimmy · Post by **swimmy** » Tue Sep 15, 2009 6:07 pm

Thanks so much.
I understand the different types of variables, but in this project there isn't really anything that was changed.
Wait, so if there are no lowercase letters in HA, then how could I tell which region of protein has more chance to mutate?
And how do I define the region of protein, by the types or where they are located in influenza? But how could I tell where they are located by the BLAST search?

Thanks,
swimmy

amyC · Post by **amyC** » Wed Sep 16, 2009 11:02 am

Hi Swimmy - I have answers for you once again.

I understand the different types of variables, but in this project there isn't really anything that was changed.

In a bioinformatics project such as Blasting the Flu, you are asking questions about differences (changes) in protein sequence. You will have to ask very focused questions or the blast output will be overwhelming.

Wait, so if there are no lowercase letters in HA, then how could I tell which region of protein has more chance to mutate?

By looking at the Blast alignments, you can see regions of the protein that are invariant, that is they never change, and regions that do vary. Regions that vary have blank spaces or +'s in the middle row of letters between the two sequences, indicating that the alignment breaks down.

And how do I define the region of protein, by the types or where they are located in influenza?

Define them by where they are located (their position by number) in the protein. If you study the protein structure (recommended) you can learn more about the function and structure of different protein domains.

But how could I tell where they are located by the BLAST search?

You can see the numbers for the amino acids in the Blast output. You may need to count from the ends. See the example below.

Query 121 YASLRSLVASSGTLEFNNESFNWTGVTQNGTSSACKRRSNNSFFSRLNWLTHLKYKYPAL 180
YASLRSLVASSGTLEFNNESF+WTGVTQNGTSSACKRRSN SFFSRLNWLTHLKYKYPAL
Sbjct 121 YASLRSLVASSGTLEFNNESFDWTGVTQNGTSSACKRRSNKSFFSRLNWLTHLKYKYPAL 180

There are changes at 142 and 161. The change at 141 is considered more conservative so a + sign is used.

If you are printing out alignments, use a font that keeps every letter the same width so the letters line up. Courier is a font that is often used.

As mentioned previously in a different post, you might want to pick a single mutation for your project to keep it focused. Then you could ask questions about how often this mutation is seen (% of all HA sequences), how does its prevalence change by year, what year did it first appear in the database, etc. You can use the mutated version of the sequence as the input in BLAST to find identical mutant sequences. You could also (more ambitious) try to see if the mutation appears independently in different populations or occurred just once and then spread.

Amy
Science Buddies

swimmy · Post by **swimmy** » Wed Sep 16, 2009 2:07 pm

Thanks. I will try this as soon as possible.

swimmy

swimmy · Post by **swimmy** » Wed Sep 16, 2009 5:27 pm

so for variables, would I need to change something in the BLAST input?

thanks,
swimmy

amyC · Post by **amyC** » Thu Sep 17, 2009 10:30 am

Hi Swimmy - I have some more information from one of our staff scientists about the "variables" in this project. I want you to try and think through the issue of identifying these.

The independent variable is the one that is changed by the scientist.

In this project, can you see what "you" selected to use with BLAST?

The dependent variable responds to changes made to the independent variable.

In this project, the dependent variable relates to the alignment.

Controlled variables are quantities that a scientist wants to remain constant.

What are you keeping the "same" for each of your searches?

See if you can use this information to come up with what the variables might be.

Amy
Science Buddies

swimmy · Post by **swimmy** » Thu Sep 17, 2009 3:53 pm

Independent: type of flu
Dependent: region where the protein mutates
Constant: Same type of BLAST search, same type of flu

Am I on the right track?
i want to compare which region of protein mutates in flu each year... or different strains. If I do each year, can I just do the most common flu that year?

swimmy · Post by **swimmy** » Fri Sep 18, 2009 10:03 pm

ok, i don't really get how you can see which number protein the mutation is. Is it 1-20 or the numbers such as 141?
also, what does more conservative mean?
what do you mean by one mutation?

swimmy · Post by **swimmy** » Sat Sep 19, 2009 2:21 pm

how would I tell where the mutation is with so many alignments?
do i just see where a mutation occurs in one alignment?
i have to turn in science fair application by tuesday, september 22.
I have to write my procedure, hypothesis, variables, make a chart/graph/data table and abstract by then.
If I compare different types of flu would I be able to make a pie chart?

swimmy · Post by **swimmy** » Sat Sep 19, 2009 4:15 pm

This is how the science fair application is like and how i think im going to enter it:

Student Participants: (I will not put my name in the forum)

Title of Project: Do flu viruses from different years change at the same regions of protein?

List your procedures/steps you'll complete:*
1) Go to the Flu Activity & Surveillance page at The U.S. Centers for Disease Control and Prevention (CDC) website: http://www.cdc.gov/flu/weekly/fluactivity.htm.

2) Click on "Go" next to the year 2008-2009 “Current Weekly Influenza Report”.

3) Read the information on the page. In particular, find the paragraph with the heading "Antigenic Characterization." It will tell you the name of the virus strains that are candidates as vaccines.

4) You can obtain the sequences for these strains from the NCBI GenBank website: http://www.ncbi.nlm.nih.gov/Genbank/.

5) Select "Protein" with the “Search” drop-down menu. Type in the name of the flu strain (use A/California/07/2009 (H1N1) in the search box.

6) Full-length HA is 566 amino acids. In order to retrieve full length-entries, add this text to the search box AND 566[SLEN]. Click "Go."

7) Click on the active link for the HA protein page.

Copy the accession number for the full-length HA protein. Click on the "BLAST Sequence" link (column on right side of page).

9) The BLAST page will pop up. The database should be set automatically to "non-redundant protein" and the algorithm should be "blastp." Click the BLAST button.

10) The BLAST report contains alignments of your search sequence against sequences in the database. The regions of the protein that is most likely to change show up as lowercase letters in the alignment.

11) Repeat Steps 1-10 with past flu seasons (2000-2008) and compare.

List the materials you will use: computer with high speed internet

Independent Variables: type of flu

Dependent Variables: region where protein is likely to mutate

Controlled Variables: same kind of BLAST search

Hypothesis:** My hypothesis is that viruses from different flu seasons do not mutate in the same regions of protein.

What research will you do before experimenting?:*** I will research BLAST (internet tool that I will be using), flu virus, flu vaccine, and the seasons of flu.

How will you display your results? (Create a table/graph/chart):****

*Please give me feedback for my hypothesis.
**In the application, I don't need to explain my hypothesis (as I was not supposed to conduct any experiment. I wanted to try this in case there was a problem...)
***Whoops... I did my research already. Have any suggestions for any other research?
****What should I make a table/graph/chart for? All I can think of is a pie chart comparing the regions of protein where the flu virus is most likely to mutate.

swimmy · Post by **swimmy** » Sat Sep 19, 2009 7:43 pm

Ok, I don't really get how I can get one mutation from all those strains of flu?

swimmy · Post by **swimmy** » Sat Sep 19, 2009 8:37 pm

If I compared mutations in types of flu by year, would it make more sense to limit it to only 2009 mutations (if I'm doing 2009) or just BLAST without specifying?

swimmy · Post by **swimmy** » Sat Sep 19, 2009 9:19 pm

The way I see it, if I do the year specified mutations, i get mutations for the flu that year, and if i don't i have to do the different types of strains.
Is this right?
I'm going to do it like this.

swimmy · Post by **swimmy** » Sat Sep 19, 2009 9:50 pm

Ok, so when I did the swine flu BLAST without specifying the year of mutations, I got the following:
Mutations by strain
H1N1 (2009)
-212 (4)
-101 (74)
-217
-333 (73)
-410 (2)
-443 (2)
-385
-166
-520
-8 (2)
-544 (2)
-467 (2)
-240 (4)
-225 (23)
-48 (3)
-476 (4)
-286 (4)
-75

The most common mutation is at 101/566

Is this how I would say the results?

And did I do the right thing by looking at the whole blast search for the mutations?

BLASTing Flu Viruses

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