Re: mammalian transgene expression - 未名空间MITBBS历史存档

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Re: mammalian transgene expression

Re: mammalian transgene expression# Biology - 生物学

c*t2003-07-14 07:07

1 楼

for a long time people already know that the presence of an intron will
signicantly increase the expression level of genes. the classic example is
beta-globin. w/o intro its cDNA barely expresses in cells. with an intron
even an irrelevant intron (insulin intron for example) it expresses at very
high level. you may wanna search with key word "intron, expression (beta-globin)
but i don't know much what's the situation in transgenic mice though.

C*n2003-07-14 07:07

2 楼

yeah apparently beta-globin transgene expression is intron-dependent,but other
genes maybe not, actually my lab and another lab in the same floor each got a
transgenic mice line with desired expression level w/o adding any introns.
I'm using the same vector pTRE as they did,maybe not that risky, should I add
an intron to my construct??

(beta-globin)
put
in
the

【在 c*****t 的大作中提到】

: for a long time people already know that the presence of an intron will
: signicantly increase the expression level of genes. the classic example is
: beta-globin. w/o intro its cDNA barely expresses in cells. with an intron
: even an irrelevant intron (insulin intron for example) it expresses at very
: high level. you may wanna search with key word "intron, expression (beta-globin)
: but i don't know much what's the situation in transgenic mice though.

c*t2003-07-14 07:07

3 楼

gee, poly(A) will contain an intron? you mean 3' UTR ba. there
are some commerical available vectors containing an intron. as
far as i know they usually put the intron at 5'UTR. coz you have
to pay attention to NMD. when you put an intron at 3'UTR it
becomes a classic example of NMD if your intron locates more than
50-55nt downstream of the stop codon.

【在 C******n 的大作中提到】

: yeah apparently beta-globin transgene expression is intron-dependent,but other
: genes maybe not, actually my lab and another lab in the same floor each got a
: transgenic mice line with desired expression level w/o adding any introns.
: I'm using the same vector pTRE as they did,maybe not that risky, should I add
: an intron to my construct??
:
: (beta-globin)
: put
: in
: the