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为了克隆黄河裸裂尻生长激素(GH)基因并获得表达产物,试验采用异硫氰酸胍法提取黄河裸裂尻脑组织总RNA,以分离的RNA为模板,采用RT-PCR扩增获得GH基因cDNA,将cDNA插入质粒pQE30中,并在大肠杆菌RB791中表达,经异丙基-β-D-硫代吡喃半乳糖苷(IPTG)诱导后,SDS-PAGE电泳检测蛋白的表达情况。结果表明:扩增后黄河裸裂尻GH基因长度为508 bp,黄河裸裂尻GH基因与青海湖裸鲤的同源性很高,电泳显示出1条新的分子质量约为14.4 ku的特异性条带。
In order to clone the GH gene and obtain the expression product, the total RNA of Brachiaria fistula was extracted by guanidinium isothiocyanate method. The isolated RNA was used as a template to obtain the GH The cDNA was inserted into plasmid pQE30 and expressed in E.coli RB791. The protein was detected by SDS-PAGE electrophoresis after induced by isopropyl-β-D-thiogalactopyranoside (IPTG). The results showed that: After amplification, the length of GH gene of Crassostreae of the Yellow River was 508 bp. The homology of the GH gene of Crassoprazole yellow tree with that of Qinghai Lake was high. One new molecular mass of 14.4 ku was identified by electrophoresis Sexual bands.