【摘要】 目的构建人紧密连接蛋白claudin-6的cDNA真核表达载体。方法应用RT-PCR、T-A克隆、限制性内切酶切及亚克隆等技术将claudin-6基因全长克隆入真核表达载体pcDNA3.1(+)质粒中。结果酶切鉴定、PCR扩增以及序列分析表明已经将claudin-6基因全长序列克隆入真核表达载体pcDNA3.1(+)质粒中。结论获得了人claudin-6基因的全长cDNA片段,构建了claudin-6原核克隆载体和真核表达载体。实验中发现,中国人claudin-6基因的461位点碱基为A,而GeneBank中西班牙人claudin-6基因的461位点碱基为G,说明claudin-6基因461位点可能存在多态性。更多还原

【Abstract】 Objective To construct human tight junction protein claudin-6 eukaryotic expression vector.Methods The cDNA fragment encoding human claudin-6 was obtained from mRNA of normal human breast tissue by using reversal transcript-polymerase chain reaction(RT-PCR).Then it was cloned into the pMD 18-T vector.After nucleotide sequencing,the cDNA was then inserted into the eukaryotic expression vector pcDNA3.1(+).Results The whole claudin-6 cDNA was cloned to eukaryotic expression vector pcDNA3.1(+) determined by enzyme digest,PCR and sequence analysis.Conclusion We get the human claudin-6 whole cDNA fragment,construct claudin-6 pronucleus cloning vector and eukaryotic expression vector. Besides we find that Chinese claudin-6 gene 461 locus base is A,but GeneBank Spanish claudin-6 gene this locus base is G,suggesting there is polymorphism on the 461 locus of claudin-6 gene.更多还原