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目的将目的基因NCAM1与质粒载体p HBAd-MCMV-GFP进行连接,得到重组质粒p HBAd-MCMV-GFPNCAM1。方法对目的基因和质粒载体分别用内切酶Not I和Nsil进行双酶切,产物回收后进行连接、转化,得到重组质粒p HBAd-MCMV-GFP-NCAM1。转化后的菌液进行PCR阳性克隆鉴定,并对阳性样本进行测序。结果首先对NCAM1进行扩增,从实验结果看符合预期效果,在双酶切、连接和转化实验后,对构建完成的重组质粒进行鉴定,PCR阳性克隆鉴定结果显示重组质粒构建成功,NCAM1已连接进入重组质粒中,测序的结果进一步印证阳性鉴定结果。结论带有目的基因NCAM1的重组质粒构建成功。
Objective To construct a recombinant plasmid p HBAd-MCMV-GFPNCAM1 by linking NCAM1 with plasmid p HBAd-MCMV-GFP. Methods The target gene and plasmid vector were digested with restriction endonucleases Not I and Nsil, respectively. The products were recovered, ligated and transformed into the recombinant plasmid p HBAd-MCMV-GFP-NCAM1. The transformed bacteria were identified by PCR positive clones and the positive samples were sequenced. Results The NCAM1 was first amplified and the results were in line with the expected results. After double digestion, ligation and transformation experiments, the constructed recombinant plasmids were identified. PCR positive clones showed that the recombinant plasmids were successfully constructed and NCAM1 was connected Into the recombinant plasmid, the sequencing results further confirm the positive identification results. Conclusion The recombinant plasmid with the target gene NCAM1 was successfully constructed.