Suggestion for carrying out experiments with the genes involved in alkalophily of Bacillus sp.

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shrinivas Dengeti - 08 Aug 2008 10:09 GMT

I have planned to carry out characterization of genes involved in
alkalophilic nature of thermoalkalophilic Bacillus sp.
Somebody can suggest me some of the methodology involved in PCR
amplification of the genes or preliminary experiments required to
understand molecular basis behind the nature.
D. Shrinivas
Research Scholar

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Bob - 09 Aug 2008 03:03 GMT

>I have planned to carry out characterization of genes involved in
>alkalophilic nature of thermoalkalophilic Bacillus sp.
>Somebody can suggest me some of the methodology involved in PCR
>amplification of the genes or preliminary experiments required to
>understand molecular basis behind the nature.

What do you already know? Do you have a genome sequence? Do you know
genes of interest? What do you know about them? Predicted protein
sequences often offer clues about function. Do you have an expression
system for Bacillus genes?

bob

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shrinivas Dengeti - 11 Aug 2008 16:20 GMT

> On Fri, 8 Aug 2008 02:09:29 -0700 (PDT), shrinivas Dengeti
>
[quoted text clipped - 11 lines]
>
> bob

i have different gene sequence of the xylanase from the Bacillus sp.
and i have PCR amplified the gene..but not yet sequenced.
I want to know whether tranport protein play any role in the
thermoalkalophilic nature or anyother factor responsible for it....if
anyone workin on the aspect.please help me in this matter...

with regards
D.shrinivas

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Bob - 14 Aug 2008 04:12 GMT

>i have different gene sequence of the xylanase from the Bacillus sp.
>and i have PCR amplified the gene..but not yet sequenced.
>I want to know whether tranport protein play any role in the
>thermoalkalophilic nature or anyother factor responsible for it....if
>anyone workin on the aspect.please help me in this matter...

A couple of quick thoughts. You've probably thought of these points
yourself, so Ill be brief.

It is interesting to have an extracellular enzyme from an alkalophile.
One can compare its properties with a "normal" relative, and then look
for an explanation in the 3D structure. Do you have a good 3D
structure for any of the related enzymes?

Of course, the general alkalophilic nature of the bug probably lies
largely in keeping the cytoplasm near "normal".

Do you know about the following papers:

http://springerlink.metapress.com/content/h3m1703x3711215t/?p=1bd8409162894aaa99
d979a1fe61676d&pi=5

http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6V24-4JRM055-1&_user=4
420&_rdoc=1&_fmt=&_orig=search&_sort=d&view=c&_version=1&_urlVersion=0&_userid=4
420&md5=6c95e774252e769ea34297f1bc792e49

I bet you already have them :-) -- but figured it safer to ask, to be
sure. If you don't have access, feel free to email me for copies, at
my yahoo acct: b_bruner

regards,

bob

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shrinivas Dengeti - 16 Aug 2008 13:31 GMT

thank you
bob..
for showing such a interest in helpin me out.
n thank you for the link too.
hey u still didnt got me..actually to say..i want to know whether the
transport involved in maintaining the alkalophilic nature of bacillus
for ex. ATPase , Na/H antiporyter pump,.. and one more thing do they
play any role in expresion of the enzyme
thank yoyu
With regards.
D.shrinivas

> >i have different gene sequence of the xylanase from the Bacillus sp.
> >and i have PCR amplified the gene..but not yet sequenced.
[quoted text clipped - 26 lines]
>
> bob

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Natural Science Forum / Biology / Microbiology / August 2008

Suggestion for carrying out experiments with the genes involved in alkalophily of Bacillus sp.