Lactic Acid Bacteria - Challenges in working with LAB culturing, cloning and plasmid (Jun/23/2011 )

Hi everyone,

I am super excited with my new project which deals with vector construction by using Lactic acid bacteria plasmid. However I have a very basic and general question for those who have worked with Lactic acid bacteria. What are the challenges and advice you would like to share with me?

Thank you in advance for your contributions, I would really appreciate any reference material that you all would recommend.

Best wishes,
It'sme

You want to know some basic microbiology skills? Or?
You never worked with bacteria before?

pito on Fri Jun 24 22:25:04 2011 said:

Hi Mr. Pito,

I have never worked with bacteria. I would certainly appreciate basic techniques/workflow in constructing plasmid with insert and using bacteria as delivery vector (carrying the plasmid/ binding to the protein expressed by the recombinant plasmid).

I would also appreciate in how to carryout immunogenecity study in inducing the immune response towards antigenic protein presented through the vector.

Thank you for your time and attention. I hope you and everyone could help in giving guidance.

Best regards,
It'sMe

It'sNotMe on Thu Jun 30 10:04:04 2011 said:

oh dear....

what you is almost impossible to answer.

Its too general, too much of the basics ..

You need to get yourself a copy of a good micrbiologybook like Brock Biology of microorganisms or prescotts book of microbiology.
Or maybe a book with standard microbiology techniques like : Microbiology Lab Manual


Anyway, its impossible to answer your questions here.. they are not specific and too diverse.
Nobody here can type an entire microbiology course here.

I think its best you start reading and when you have specific questions you can ask them.

Dear Mr. Pito,

Thank you for your reply and reference suggestions. I am indeed reading books like sambrook and prescot. I am just trying to find out tips and practically optimised work flow followed by experienced people (especially those who have worked with lactic acid bacteria) . Well I guess may be I am too early into my research to ask questions. Thanks again Mr. Pito I will certainly checkout those references you have named.

Best wishes,
It'sMe

It'sNotMe on Thu Jun 30 14:25:08 2011 said:

Its too early... You havent done anything yet.. so you havent really faced any problems...
Just start by training to understand the basic work, how to work with bacteria etc.. try to get as many information as possible on working with bacteria in general, techniques etc.. and then you can look up some papers about lactic acid bacteria and then you can start figuring out how to do it practically...

But to be honest: I do not understand it that you need to do this without any help/supervision from someone with experience in the field...

Dear Mr. Pito,

Thank you for your valuable advice again. I do have a supervisor and a co-researched, PhD candidate. The latter is back in his country for semester break so cant really communicate. Anyway, I thin I am too nervous and rushing into stuff. I am doing heavy reading now on cloning, cell culture, expression vector, immunogenecity and etc. Thank you, and sorry for troubling you with silly questions. I'll be back with specific question later on.

Best wishes,
It's Me

It'sNotMe on Fri Jul 1 08:14:25 2011 said:

its best to get a general idea about what you are trying to do.. try to understand the basics like for example "cutting" a piece of DNA from 1 bacterium and inserting this in a plasmid and then transferring the plasmid in another bacterium to grow it... etc..
Just the simple basic stuff, you dont need to know all the details yet.