conjugation

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conjugation

Postby kamranban » Thu Oct 22, 2009 11:19 pm

Hi
I have a microbiology project and I need some help.
my project is based on the danger of antibiotics to the normal flora (esp. the normal flora of gut).
we all know that when we use antibiotics not only the pathogens but also the normal flora are destroyed. how good life could be if only the pathogens were destroyed by antibiotics. we can make bacteria of normal flora resistant to antibiotics (at present, only one bacteria, and only one antibiotic), and then add that resistant bacteria to the normal flora. but there is a problem here and that is conjugation. for example; we want to make E.coli resistant to an antibiotic. then if E-coli 0157:H7 enters our body, the E.coli of our normal flora conjugates with E-coli 0157:H7 and it becomes resistant to that kind of antibiotic too.

what I thought I could do to solve this problem was:
1) cutting down the conjugative pili.
2) finding a bacteria in normal flora that doesn’t conjugate.
3) finding a bacteria in normal flora that doesn’t have a pathogen family.
are these ways possible or what are the other ways that I can do? :?:
kamranban
 
Posts: 10
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Occupation: student 10th grade
Project Question: antibiotic resistance and normal flora
Project Due Date: 1 year
Project Status: I am conducting my research

Re: conjugation

Postby barretttomlinson » Fri Oct 23, 2009 1:14 pm

Hi,

If you have not read this project writeup I think you will find it very useful. Pay particular attention to the bibliography links:

http://www.sciencebuddies.org/science-f ... p013.shtml

You may also find these sites informative:

http://www.bio-medicine.org/biology-def ... njugation/

http://www.bookrags.com/Bacterial_conjugation

http://www.mun.ca/biochem/courses/3107/ ... ation.html

http://en.wikipedia.org/wiki/Bacterial_conjugation

http://www.slic2.wsu.edu:82/hurlbert/mi ... lab14.html

http://mbe.oxfordjournals.org/cgi/reprint/15/4/385.pdf

I am not an expert in this field, so I hope others will answer your more specific questions.
However I think these resources will get you started on thinking through your experimental plan.

Good luck! It sounds like a fascinating project!!!

Best regards,

Barrett L. Tomlinson
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Re: conjugation

Postby barretttomlinson » Sat Oct 24, 2009 2:07 pm

Hi,

This paper on interspecies transfer of plasmids should be of great interest to you:

http://mbe.oxfordjournals.org/cgi/reprint/15/4/385.pdf

In considering your project question you really need to be aware of three different mechanisms of gene exchange: transformation, transduction, and conjugation:

http://www.emunix.emich.edu/~rwinning/g ... actrec.htm

http://www.biologyreference.com/Ar-Bi/B ... etics.html

The more I have looked into the questions you posed in your post the more interesting it looks.
I think you have the seeds of ideas for really interesting project. I wish you the best of luck with it!

Best regards,

Barrett L. Tomlinson
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Re: conjugation

Postby barretttomlinson » Sun Oct 25, 2009 1:00 pm

Hi,

Your post asked about possible ways to avoid antibiotic resistance in pathogens while immunizing beneficial bacteria against the antibiotics by using conjugation. My take on your question is that the essence of your idea is combating antibiotic resistance in pathogens - the protections of beneficial bacteria is desirable,but of secondary importance. The idea of modifying or eliminating pilli from the beneficial bacteria sounds like a long shot to me, as many resources I have seen and cited to you inicate pilli are essentially involved in conjugation, and interspecies conjugation appears widespread. I would suggest for your creative thinking you consider ways to reverse antibiotic resistance, perhaps by trying to use conjugation to insert an antisense RNA gene to the antibiotic resistance gene into an F+ plasmid in the beneficial bacteria. This might enable the beneficial bacteria to disarm the pathogens by conjugation.

Some backgound on antisense RNA in this area:
Read this first article if you read nothing else. It is a good overview of the use of antisense RNAs to fight antibiotic resistance!

http://jac.oxfordjournals.org/cgi/content/full/dkn467v1

This paper targets your interest almost exactly, as I understand it:

http://www.ncbi.nlm.nih.gov/pmc/article ... 1-0636.pdf

http://jb.asm.org/cgi/content/abstract/177/7/1655

http://jb.asm.org/cgi/content/abstract/178/1/306

http://jb.asm.org/cgi/content/abstract/175/4/1026

http://aac.asm.org/cgi/content/full/47/11/3485


http://jb.asm.org/cgi/content/full/182/ ... alsMethods

http://www.ibiblio.org/ecolandtech/Soil ... 00838.html

http://www.bioline.org.br/request?oc03093


I think this book could be very useful to you:

http://books.google.com/books?id=9Hw-ms ... ce&f=false

Please note that the above site will help you locate the book in a library close to you!

Thank you for asking the question you did. I have had a lot of fun researching this subject, and learned a lot myself. I think you have found an outstandingly interesting subject to study. I wish you a very successful project!!!!!

Best regards,
Barrett L. Tomlinson
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Re: conjugation

Postby kamranban » Tue Oct 27, 2009 7:43 am

Hi
Thank you very much.
The links in your first post were really useful (because I have not read your new links yet)
I found my answer in that articles. But it has a problem. The solution : We can only put f- bacteria in the gut ,because the f- bacteria is not donor and its only recipient and the connection between male and female is only one way connection (from male to female). But the problem is that when another bacteria comes into the gut and do a conjugation with the normal flora bacteria they send their f-factors and then our female bacteria changes into the male one.
And I think that you misinformed about a part of my project I don’t want to make bacteria resistant to an antibiotic by using conjugation. I want to select some resisted bacteria from the culture and conjugation is a disease in my project. At present I want to find out a way to stop conjugation or not letting the bacteria be conjugated.
thanks kamran :D
Quis custodiet ipsos custodes?
kamranban
 
Posts: 10
Joined: Wed Oct 21, 2009 11:43 pm
Occupation: student 10th grade
Project Question: antibiotic resistance and normal flora
Project Due Date: 1 year
Project Status: I am conducting my research

Re: conjugation

Postby aelin » Tue Oct 27, 2009 5:57 pm

Hi Kamran,

I haven't had a ton of experience with bacterial conjugation, but the question you raise is an interesting ones.

First, a few potential roadblocks that I spot. It seems to me that you are mostly focusing on gastrointestinal conjugation (several groups of bacteria will never conjugate with gut flora), and as mentioned above, in such cases where strains like O157:H7 are present, antibiotics to clear out the virulent bacteria are generally preferred even though they are still toxic to the normal flora. O157:H7, along with a few other strains, is particularly nasty because it produces Shigella-like toxins that selectively target the larger ribosomal subunit of our eukaryotic cells (interestingly enough, this is the reverse of several of our antibiotics--many antibiotics work by blocking the prokaryotic 30S ribosomal subunit, preventing the bacteria from producing proteins and ultimately leading to cell death; imaging the 30S subunit was actually just recently awarded a Nobel prize).

It further seems that your project would be limited even in the types of gastrointestinal bacteria it deals with. Certain bacteria like cholera actually produce toxins that trigger the body to wash out the normal flora, and in such a case, antibiotics would definitely produce more good than harm.

Nevertheless, this actually makes it easier for you to do a project at school since it seems like the simplest bacteria to work with would be E. coli. This article posted above (http://www.ncbi.nlm.nih.gov/pmc/article ... 1-0636.pdf) seems promising in blocking conjugation--be careful though, this seems to mostly be dealing with the donor bacteria, not the recipients that you want to target, but is still a start in terms of understanding the genes involved in conjugation.

Another route to blocking conjugation may be to get the bacteria to produce and secrete an enzyme that catalyzes the F factor (I'm not sure this is known, I'm looking into it, but imaginably such an enzyme exists).

Also, another thought off of the top of my head is the maping of E. coli genome. I know that to do so, they used bacteria that were especially selected for high frequency conjugation to initiate conguation, and they then interrupted it via a blender or some kind of force. Obviously that would not be a valid strategy inside of the gut, but looking into the specific mechanism of interruption (eg force, disruption of certain enzymes) may provide some clues as to more ways to interrupt conjugation.

Hope this helps,
Aaron Lin
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Re: conjugation

Postby kamranban » Wed Oct 28, 2009 11:18 am

Hi Aaron
When I exampled E.coli of normal flora and O157:H7 I knew that they must be from a same family (e.g. both are E.coli)
We have 2 kinds of antibiotics: 1.specialty antibiotics 2. Non specialty Antibiotics. The first one doesn’t have any disadvantages but the second one not only kills the pathogens but also it kills the bacteria of normal flora , I want to make antibiotics more beneficial and less harmful. (Sort of making a 2nd type antibiotic, half specialty)
Thanks kamran
Quis custodiet ipsos custodes?
kamranban
 
Posts: 10
Joined: Wed Oct 21, 2009 11:43 pm
Occupation: student 10th grade
Project Question: antibiotic resistance and normal flora
Project Due Date: 1 year
Project Status: I am conducting my research

Re: conjugation

Postby kamranban » Wed Nov 04, 2009 9:39 am

Hi everyone
Thanks for your links they are very useful for such a project and any one who wants to have project in this case must read that articles.
Now 1.I know how to stop conjugation in a culture. And 2.I know how to stop f+ bacteria from conjugating. Actually I can use the first way only in lab (not in the gut). And going throw the second way can’t help me because there are always some bacteria that come into the gut from outside and I can’t stop their conjugation with our normal flora , What I should do is stopping f- bacteria from being conjugated by others. Any Idea?
Thanks kamran banan
Last edited by kamranban on Sat Nov 07, 2009 5:31 am, edited 1 time in total.
Quis custodiet ipsos custodes?
kamranban
 
Posts: 10
Joined: Wed Oct 21, 2009 11:43 pm
Occupation: student 10th grade
Project Question: antibiotic resistance and normal flora
Project Due Date: 1 year
Project Status: I am conducting my research

Re: conjugation

Postby barretttomlinson » Wed Nov 04, 2009 11:09 pm

Hi,

Here is a report of an inhibitor to bacterial conjugation:

http://mic.sgmjournals.org/cgi/content/ ... 51/11/3517

Here is a patent on the subject:

http://www.freshpatents.com/-dt20090903 ... escription

And here are papers on the same idea:

http://virtualgenomeproject.blogspot.co ... tance.html

http://www.pnas.org/content/104/30/12282.full.pdf+html

Hope this is what you needed.

Good luck!

Barrett L. Tomlinson
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Re: conjugation

Postby kamranban » Sat Nov 07, 2009 5:39 am

barretttomlinson wrote:Here is a report of an inhibitor to bacterial conjugation:

http://mic.sgmjournals.org/cgi/content/ ... 51/11/3517




Hi
I really appreciate you guys. that links were very useful(mostly the link above). now sth. about 80% of theoric part of my project is finished.
best wishes
kamran banan
Quis custodiet ipsos custodes?
kamranban
 
Posts: 10
Joined: Wed Oct 21, 2009 11:43 pm
Occupation: student 10th grade
Project Question: antibiotic resistance and normal flora
Project Due Date: 1 year
Project Status: I am conducting my research

Re: conjugation

Postby kamranban » Wed Nov 11, 2009 8:41 am

Hi everyone
my mentor told me :cry: that I can't stop :cry: conjugation in gut :cry: by using fatty acids.
(the link in the post above)
I didn’t understood his reason. :cry: and he wants to change my project a bit :cry: but I strongly believe in my own project. :? How can I find a university professor to ask my question from him? and how should I start being in contact with them?
thanks
kamran banan
Quis custodiet ipsos custodes?
kamranban
 
Posts: 10
Joined: Wed Oct 21, 2009 11:43 pm
Occupation: student 10th grade
Project Question: antibiotic resistance and normal flora
Project Due Date: 1 year
Project Status: I am conducting my research

Re: conjugation

Postby ladan mz » Thu Nov 12, 2009 12:52 pm

WhY my reply has been deleted????!!!!! :?: 8)
Anyway, http://www.ncbi.nlm.nih.gov/sites/entrez
check this page, it could be help you.
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