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XUMIN6891金虫 (初入文坛)
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[求助]
把这段中文翻译成英语
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| 摘要：目的：构建抗菌肽LNK-16基因6拷贝，12拷贝串联体，并将目的基因串联产物构建到表达载体pET30a(+)上。方法：以实验室构建好的LNK-16基因3拷贝的重组质粒为模板设计引物，通过PCR扩增合成2段3拷贝的LNK-16片段，分别命名为G1，G2。其中片段G1的上下游分别设有Nco I和Xho I酶切位点，片段G2的上下游分别设有EcoR I和Nco I酶切位点，分别把G1和G2双酶切回收，然后将pET30a(+)载体用EcoR I ，Xho I双酶切回收，最后在一个体系里将片段G1，G2，和pET30a(+)载体连接。这样得到含6拷贝的LNK-16基因。然后再以它为模板设计引物，通过PCR扩增合成2段6拷贝的LNK-16片段分别命名为G3，G4。其中G3的上下游分别设有EcoR I和Hind III酶切位点，G4的上下游分别设有Hind III和Xho I酶切位点，分别把G3和G4双酶切回收，然后将G3，G4和经过EcoR I 、Xho I双酶切回收的pET30a(+)载体连接，这样就得到含12拷贝的LNK-16基因。结果：PCR鉴定、双酶切鉴定和DNA测序证明6拷贝，12拷贝基因的重组质粒构建成功。结论：该方法能方便高效地获得所需要的多拷贝基因，为进一步克隆到表达载体进行高效表达打下基础。 |
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AnnF
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XUMIN6891: 金币+2, 翻译EPI+1, 你用机器直译的把？ 2012-05-24 17:27:36
爱与雨下: 翻译EPI-1 2012-05-24 21:39:54
XUMIN6891: 金币+2, 翻译EPI+1, 你用机器直译的把？ 2012-05-24 17:27:36
爱与雨下: 翻译EPI-1 2012-05-24 21:39:54
| Abstract: Objective: To build the antimicrobial peptide LNK-16 gene 6 copy, 12 copies of the tandem body, and the target gene series products built into the expression vector pET30a (). The method: to build a good laboratory LNK-16 gene copies of the recombinant plasmid as a template design primers synthesized by PCR amplification of paragraph 2 of 3 copies of LNK-16 fragment, were named G1, G2. Fragment G1 of the upstream and downstream, respectively, with Nco I and Xho I sites upstream and downstream of the fragment of G2 with EcoR I and Nco I restriction sites, respectively, G1 and G2, double digestion recovery, and pET30a () vector using the EcoR I and Xho I double digestion recovery, the last in a system where the fragments G1, G2, and and pET30a () vector. This LNK-16 containing six copies of the gene. LNK-16 fragments and then using it as the template design primers synthesized by PCR amplification of paragraph 2 of 6 copies were named as G3, G4, Which the G3 upstream and downstream, respectively, with EcoR I and Hind III restriction sites, the G4 upstream and downstream, respectively, with Hind III and Xho I sites, respectively, G3 and G4 double digestion recovery, then to G3, G4, after the EcoR I, Xho I double digestion recovery of pET30a () vector, so that the LNK-16 gene containing 12 copies. Results: PCR identification of restriction enzyme digestion and DNA sequencing to prove that six copies of the 12 copies of recombinant plasmid was successfully constructed. Conclusion: This method can be easily and efficiently obtain the required multi-copy gene for efficient expression lay the foundation for further cloned into the expression vector. |
2楼2012-05-24 11:04:53
