山蛭基因组DNA的提取及其PCR扩增产物的分析

Extracting of Hainan Leech Genomic DNA and its Analysis of PCR Products

利用PCR技术从海南山蛭体内分离山蛭素 (抗凝血蛋白 )基因 ,首先需获得不带有山蛭体表色素且完整的山蛭基因组DNA ,本试验通过结合使用SDS-蛋白酶裂解法和CTAB法 ,有效的去除了山蛭基因组DNA提取过程中难以去除的色素 ,得到的基因组DNA保持完整 ,无降解。以之作为模板 ,进行PCR扩增 ,获得一个长度约为 2 37bp的特异性片段 ,此片段与印度山蛭蛭素的cDNA大小几乎一致。初步表明 ,这个序列没有内含子 。

The extracting of genomic DNA is the key point to obtain the haemadin gene from Haemadipsa hainana (HHL) by PCR In this experiment,an improved method of SDS-protease combined with CTAB was used to gain the genomic DNA from HHL A specific DNA fragment about 237 bp length was acquired from PCR in which the genomic DNA served as template with a pair of specific primers The specific DNA fragment is similar to the cDNA sequence from Indian leech ( Haemadipasa sylvestris Blanchard) It suggests that the fragment is the coding sequence of HHL haemadin