发信人: adeno (浮沉), 信区: Biology
标 题: Re: help me!!
发信站: The unknown SPACE (Sun Sep 17 16:55:59 2000) WWW-POST


The method to make transgenic mice is by microinjection of
your construct DNA into the pronucleus of the fertilized
oocyte. Your construct will integrate into the chromosome
randomly, and usually tandemly in a single chromosome. The
amount of expression is dependent on the copy numbers of
your favorite gene, and also the promoter you choose

Depending on the purpose of your research, you can choose
tissue-specific or ubiquitous promoter. Sometimes, you just
want to use its own promoter. In your case, you have to use
tissue specific ones. Briefly, if a gene is exclusively
expressed in a tissue, its promoter is specific for this
tissue. For example, if you want to overexpress a protein in
adult heart ventricle, you can use alpha-myosin heavy chain;
if you want to overexpress it in embryonic heart venticle,
you should use beta myosin heavy chain promotor. (so they
are not only tissue specific but also development stage
specific)

Making constucts is easy, just ligate them together, I am
sure you know that:)




【 在 gardenia (怅然若失的栀子花) 的大作中提到: 】
:
: Thank you , but can you explain in detail?\
:
: Such as what tissue specific promoter's nature and how to do
: the insert of such promoter
:
: Thanks a lot
: 【 在 adeno (浮沉) 的大作中提到: 】
: : 【 在 gardenia (怅然若失的栀子花) 的大作中提到: 】
: : : just ask something theogically...
: : : Now that we know that a certain is only expressed in mice brain,
: : : and we need to over-express it in mice brain and other tissue as
: : : well so as to do some research on its effect, what technique can
: : : we use?
: : : Thanks a lot!!
: : put your gene after your tissue specific promoter and make
: : transgenic mice
:
:

--
母亲接过孩子搂在怀里。她瞅着他，又惭愧又欢喜地笑了笑：

“噢，我的小乖乖，你多难看，你多难看，我多疼你！”



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