大鼠钠尿肽B受体cDNA探针克隆载体的构建

Construction of cDNA probe clone vector for natriuretic peptide B receptor

目的 :构建大鼠钠尿肽 B受体 c DNA探针克隆载体 ,为研究该受体基因表达奠定基础。方法 :设计钠尿肽 B受体特异的 PCR引物 ,提取大鼠肺组织总 RNA,进行反转录 PCR,扩增目的片段 ,回收目的片段 ,将其克隆入p GEM- T载体 ,酶切鉴定并用双脱氧法测序。结果 :PCR反应产物经琼脂糖凝胶电泳鉴定与所设计引物大小一致 ,克隆入 p GEM- T载体后酶切可切出所需片段 ,测序证实为所设计序列。结论 :构建的 p GEM- T重组质粒为进一步研究该受体基因表达。

AIM: To construction of cDNA probe clone vector for B receptor of natriuretic peptide. METHODS: SD rat lungs were employed to clone the fragment of natriuretic peptide B receptor. Total RNA molecules were isolated from fresh lungs and mRNA were reversely transcriped into cDNAs. After amplification by PCR,the fragment of DNA were cloned into vector pGEM T. The method of dideoxy termination was employed to sequence. RESULT: The gene fragment of B receptor of natriuretic peptide was successfully cloned in pGEM T vector and the sequence was correct. CONCLUSION: The clone vector constructed can be a source of cDNA probe for natriuretic peptide B receptor and was useful in study its gene express.