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将新近发现的人泛素缀合酶样蛋白基因h MMS2的 c DNA特定反向克隆到本室改建的真核细胞表达载体 p MAMneo- amp-中 ,构建了表达h MMS2反义 RNA的重组质粒 .将此反义表达重组质粒 (p MAM- anti- h MMS2 )用改良的磷酸钙法转染人羊膜 FL细胞 ,建立 FL- h MMS2 -细胞系 .比较该细胞系及 FL细胞和转染了空载体 p MAMneo-amp-的 FL细胞 (FL- MAMneo细胞 )的生长速度 ,发现 FL- h MMS2 -细胞系在地塞米松诱导下h MMS2基因表达被反义抑制后导致细胞生长受阻 ,提示 h MMS2基因为细胞生长所必需 .
The newly discovered human ubiquitin conjugate-like protein gene h MMS2 c DNA specific reverse cloning into the eukaryotic cell expression plasmid pMAMneo-amp-, to construct a recombinant plasmid expressing h MMS2 antisense RNA This antisense recombinant plasmid (pMAM-anti-h MMS2) was transfected into human amnion FL cells using a modified calcium phosphate method to establish the FL-h MMS2 - cell line.This cell line and FL cells were compared and transfected Growth of FL cells (FL-MAMneo cells) in empty vector pMAMneo-amp-FL and found that h MMS2 gene expression in FL-h MMS2 - cell lines induced by dexamethasone was inhibited by antisense, resulting in cell growth retardation, h The MMS2 gene is necessary for cell growth.