【摘要】 目的　获得中国人的肺表面活性物质结合蛋白A(SP A) 6AcDNA基因克隆 ,并进行序列分析 ,为进一步研究中国人SP A基因型特点提供方法和资料 ,同时也为SP A基因表达调控。方法以正常人肺组织mRNA为模板 ,采用逆转录反应 (RT) PCR、巢式 PCR及基因克隆技术 ,获得pUC19/SP A 6A克隆 ,并测序证实。结果　从正常中国人肺组织中提取到完整mRNA ,经RT PCR、巢式 PCR扩增 ,得到约 840bp目的片段 ,分别用BamHI、HindIII和PstI酶切 ,酶切片段与分析结果一致 ,分别为 138、2 75、416、388和 417bp。将 840bp目的片段与测序载体连接 ,测序结果显示 ,与已发表的SP A6A序列相比 ,有若干个碱基不同 ,随机重复 2个克隆 ,证实位于第 19、6 2、133位密码子的改变是稳定存在的。结论　证实获得的中国人SP A基因克隆为 6A3 型。更多还原

【Abstract】 Objective To obtain and sequence Chinese cDNA clone of pulmonary surfactant protein A (SP A) in order to further study SP A gene structure, expression regulation and genetic engineering in Chinese. Methods The mRNA from normal lungs was used as template. By using the techniques of RT PCR, nest PCR and cloning, the pUC 19/SP A 6 A clone was established and sequenced. Results The high quality mRNA was extracted from the normal Chinese lung tissues and a 840 bp fragment was obtained by RT PCR and nest PCR. With BamHI, HindIII and PstI, fragments of 138 bp, 275 bp, 416 bp, 388 bp and 417 bp cDNA were obtained, which are similar to PCGENE. The 840 bp cDNA was linked with pUC 19 and sequenced. Compared with the published SP A 6A sequences, there were several bases changed, 2 clones repeated. The changes at codon 19,62 and 133 were stable. Conclusion It was approved SP A clone in Chinese was 6A 3 subtype.更多还原