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目的:从人胎盘组织克隆WNT6的全部编码区,测定并分析其基因序列。方法:利用RT-PCR方法,扩增人WNT6基因片段,将基因片段插入pMD18-T载体,限制性内切酶酶切鉴定,测定序列。结果:重组质粒pMD18-T-WNT6酶切鉴定正确,测序结果经Blast分析与GeneBank公布的WNT6基因编码区一致。结论:成功克隆人WNT6编码区基因,为体外表达其活性蛋白奠定了基础。
OBJECTIVE: To clone the entire coding region of WNT6 from human placenta and determine its gene sequence. Methods: The human WNT6 gene fragment was amplified by RT-PCR. The gene fragment was inserted into pMD18-T vector and restriction endonuclease digestion was performed to determine the sequence. Results: The recombinant plasmid pMD18-T-WNT6 was identified by restriction enzyme digestion. The results of Blast analysis were consistent with the WNT6 gene coding region published by GeneBank. Conclusion: The successful cloning of human WNT6 coding region gene laid the foundation for the expression of its active protein in vitro.