摘要
本文用含有人工合成28肽心钠素基因质粒的大肠杆菌(TAP106-pYX40),分别进行温度诱导和丝裂霉素C诱导,提取菌体中的总RNA,与γ-^(32)P标记的心钠素基因片段进行RNA打点杂交,并通过测定杂交阳性信号的总灰度值(Totalgray value),建立了人心钠素基因特异mRNA相对定量方法,从而观察到温度诱导对P_L启动子控制下的心钠素基团转录发生得慢,特异的mRAN产量较低,但转录所持续的时间长,而丝裂霉素C诱导对P_L启动子控制下的心钠素基因转录发生得快,特异mRNA产量高,但持续的时间短。
The kinetics of specific mRNA transcription from a synthetic human atrial natriuretic polypeptide (hANP) gene has been analyzed. The total RNAs were extracted from the culture of E. coli TAP106 harboring with plasmid pYX40, which contains the gene under the control of PL promoter, at various time after temperature (42℃) or mitomycin C induction respectively. Dot blot hybridization was performed using a gene fragment labelled by γ-32P-ATP as probe. The relative amount of hANP mRNA were estimated from the total grey value of each hybridization spot and the kinetic change of the mRNA synthesis was observed. The results showed that the transcription of the gene after mitomycin C induction was faster in speed, higher in level and maintained for shorter time as comparared with those after temperature induction.
关键词
Pl启动子
心钠素基因
基因转录
Human atrial natriuretic
Polypeptide gene
PL promoter
Mitomycin C
Hybridization
