Gnrh / trs gene was cloned into the pmd18 - t vector , the recombinant pycl plasmid was identified and analyzed by corresponding restriction endonuclease and nucleotid sequence , lt is indicated that the gnrh / trs gene has been cloned successfully , then this gene was cloned into the expression vector pgex - 6p - l , the recombinant pyc2 plasmid was identified and analyzed by corresponding restriction endonuclease , it is indicated that the gnrh / trs gene has been cloned successfully . the recombinant plasmid pyc2 was transformed into the bl21 ( ed3 ) plyss prokaryotic expression system 将其克隆到pmd18 - t载体上,构建重组质粒pyc1 ,经酶切鉴定筛选出阳性克隆pyc1 ,进一步的序列分析确认其中gnrh trs序列的正确性。 pyc1经bamhi和ecori酶切得到gnrh trs片段,然后将此片段克隆到表达载体pgex - 6p - 1上,构建重组质粒pyc2 。经酶切鉴定筛选出阳性克隆pyc2 ,并转化到大肠杆菌bl21 ( ed3 ) plys实现原核表达。