人DNMT 3B7真核表达载体的构建和鉴定

Construction and identification of eukaryotic expression plasmid human DNMT 3B7

目的构建人DNMT3B7真核表达载体pCMV-DNMT3B7。方法据Genebank中人DNMT3B7cDNA序列设计并合成特异性引物,以人DNMT3B1为模板,利用PCR技术扩增出DNMT3B7,并将扩增产物克隆到真核表达载体pCMV-2B中。结果 PCR扩增得到人DNMT3B7的cDNA片段大小为1.1kb,重组子利用限制性内切酶进行酶切、PCR鉴定和DNA序列分析得到真核表达载体pCMV-DNMT3B7。结论成功构建了人DNMT3B7真核表达载体,为进一步研究DNMT3B异构体提供了条件。

Objective：To construct and identify the eukaryotic expression plasmid for human pCMV-2B-DNMT 3B7.Methods： According to the published human cDNA sequence in Genebank,a pair of primers were respectively designed and synthesized.pCMV-2B-DNMT 3B1 was used as template.After amplification with polymerase chain reaction（PCR）,the product was cloned into pCMV-2B vector.The recombinants were finally sequenced and identified by restrictive endonuclease digestion.Results： pCMV-DNMT3B7 eukaryotic expression vectors was successfully constructed,and it was identified by PCR,double restrictive endonuclease digestion and sequence analysis.Conclusion： The DNMT 3B7 eukaryotic expression vectors are successfully constructed and identified.