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[ÇóÖú] PET-HMÊǸöʲôÖÊÁ£°¡£¬ÇóÖúÏà¹ØÐÅÏ¢ ÒÑÓÐ1È˲ÎÓë

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ÎÄÏ×Ô­ÎÄ£ºThe expression
plasmid PET-HM was introduced into the T7-RNA-polymerasecontaining
host E. coli BL21(DE3) (Studier et al., 1990). T7
RNA polymerase is used to drive the expression of DNA located
downstream of the T7 promoter of the PET vector. Cells harboring
PET-HM or PET-3a (control) were each grown under agitation
at 37¡ãC in Luria-Bertani medium containing 100 pg carbenicillidml,
in the presence or absence of 100 pM lipoic acid,
to an A,,, of 0.5 (about 2-2.5 h). Induction of the target prcitein
was started by addition of isopropylthio-b-D-galactopyranclside
to 0.4 mM. To monitor the induction of H protein 1 ml of culture
was sampled at various times after induction. The cells were
centrifuged (14000Xg, 1 min) and suspended in 100 p1 5DS/
PAGE sample buffer (Laemmli, 1970). After heating at 100¡ãC
for 3 min, 10 p1 of the sample (corresponding to 100 pl of culture)
were subjected to
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jie123400

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soloaaaa: ½ð±Ò+5, ¡ï¡ï¡ï¡ï¡ï×î¼Ñ´ð°¸ 2016-08-09 14:21:40
xn8008: ½ð±Ò+1, »¶Ó­·¢Ìû½»Á÷ 2016-08-10 07:24:40
Â¥Ö÷£¬ÄãÔÙ°ÑÎÄÏ׺úÿ´Ò»¿´PET-HMÓ¦¸Ã¾ÍÊÇPET-3a×°ÁËÄ¿µÄ»ùÒòµÄ±í´ïÔØÌ壬֨бàÁ˸öÃû×Ö¡£

Ô­ÎÄÊDz»ÊÇÕâ¸ö£º¡°The Ndel-BamH1 fragment was subcloned into the
expression vector PET-3a (Stratagene), digested with Ndel and
BamHl , to yield the expression plasmid PET-HM.¡±
2Â¥2016-08-09 13:53:45
ÒÑÔÄ   »Ø¸´´ËÂ¥   ¹Ø×¢TA ¸øTA·¢ÏûÏ¢ ËÍTAºì»¨ TAµÄ»ØÌû

soloaaaa

ľ³æ (ÕýʽдÊÖ)

¡ï
xn8008: ½ð±Ò+1, »¶Ó­·¢Ìû½»Á÷ 2016-08-10 07:24:46
ÒýÓûØÌû:
2Â¥: Originally posted by jie123400 at 2016-08-09 13:53:45
Â¥Ö÷£¬ÄãÔÙ°ÑÎÄÏ׺úÿ´Ò»¿´PET-HMÓ¦¸Ã¾ÍÊÇPET-3a×°ÁËÄ¿µÄ»ùÒòµÄ±í´ïÔØÌ壬֨бàÁ˸öÃû×Ö¡£

Ô­ÎÄÊDz»ÊÇÕâ¸ö£º¡°The Ndel-BamH1 fragment was subcloned into the
expression vector PET-3a (Stratagene), dig ...

ÊÇÎÒ´íÁË£¬Ã»×Ðϸ¿´~ ллÀ²~ ÄãÒ²×öÕâ¸öÂð£¿
3Â¥2016-08-09 14:22:45
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jie123400

Í­³æ (СÓÐÃûÆø)

¡ï
xn8008: ½ð±Ò+1, »¶Ó­·¢Ìû½»Á÷ 2016-08-10 07:24:53
ÒýÓûØÌû:
3Â¥: Originally posted by soloaaaa at 2016-08-09 14:22:45
ÊÇÎÒ´íÁË£¬Ã»×Ðϸ¿´~ ллÀ²~ ÄãÒ²×öÕâ¸öÂð£¿...

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