【摘要】 以海南山蛭(Haemadipsahainana)基因组DNA为模板,以合成的2段30个寡聚核苷酸的序列为引物,经过PCR扩增,分离海南山蛭中的蛭素基因,获得了1条大小约237 bp的DNA片段。将其克隆到pGEM-Teasy载体中,经筛选与检测,并进行序列分析。结果显示,所克隆的海南山蛭蛭素基因的大小为231 bp,与Dr. Burkhard报道的印度山蛭蛭素cDNA序列相比,其核苷酸的同源性为99.9%。更多还原

【Abstract】 A specific DNA fragment about 237 bp was obtained from PCR in which the genomic DNA from Haemadipsa hainana served as template and two short specific sequences of DNA as primers. By electro phoresis on low melting agrose gel, the specific DNA fragment was retrieved, purified and then linked into pGEM-Teasy vector by linkase T4DNA. The recombinant plasmid was transfered into Escherichia coli DH5α. The transformer was first screened by IPTG/X-gal, then authenticated by PCR and by SacⅠ and ApaⅠ restriction enzyme digestion. The bacteria containing the recombinant plasmid were quickly proliferated, and the heterogeneous gene was magnified. Purified recombinant plasmid was sequenced. The result showed that the sequence of haemadin gene had a 99.9% homology with cDNA sequence of Haemadin from an Indian leech.更多还原