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PET-HMÊǸöʲôÖÊÁ£°¡£¬ÇóÖúÏà¹ØÐÅÏ¢ ÒÑÓÐ1È˲ÎÓë
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¿´ÎÄÏ×¼ûµ½Á˸öPET-HMÖÊÁ££¬²é²»µ½Ïà¹ØµÄÐÅÏ¢£¬Çó½Ì¡£ 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
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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
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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
soloaaaa
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3Â¥2016-08-09 14:22:45
jie123400
ͳæ (СÓÐÃûÆø)
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- ×¢²á: 2008-05-16
- רҵ: µ°°×ÖÊ×éѧ
4Â¥2016-08-09 14:41:10













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