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生物学四句翻译
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| 以牛的抗菌肽（Indolicidin）氨基酸序列为模板，在多肽C端添加Kallikrein蛋白酶切位点设计出新的抗菌肽NK-16检测出抗菌活性。根据氨基酸序列和E.coli偏爱密码子，反向翻译设计并合成四个重叠基因片段，在5’端和3’端分别导入用于表达载体构建的限制性内切位点Xho I和Nco I。利用Overlapping PCR技术，连接合成片段，将连接好的核苷酸序列克隆入pET 30a(+)载体中构建多重载体PET30a(+)-NK-16，经双酶切和PCR鉴定表明多重表达载体PET30a(+)-NK-16构建成功。多重表达载体PET30a(+)-NK-16的正确构建，为抗菌肽的原核高效表达奠定了基础。 |
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ringzhu 
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fbook(金币+100, 翻译EPI+1): 2010-09-30 18:04:39
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To detect antimicrobial activity,we take amino acid sequence of Bovine antimicrobial peptide (Indolicidin) as a template, adding Kallikrein protease cleavage sites to the C terminal of peptide to get new antimicrobial peptide NK-16. Based on amino acid sequence and E.coli preferred codons,four overlapping fragments were backward translated ,designed and synthesized. The restriction sites Xho I and Nco I which were used to express the contructino of vector were introduced onto 5 'and 3'terminal. Overlapping PCR technique was used to connect synthetic fragment and made the connected nucleotide sequence cloned into pET 30a (+) vector to construct multi-carrier PET30a (+)-NK-16.the multiple expression vector PET30a (+)-NK-16 was proved to be constructed successfully by the identification by Double digestion and PCR. the right construction of multiple expression vector PET30a (+)-NK-16 lay the groundwork for the highl expression of prokaryotic antimicrobial peptides. |
2楼2010-09-30 11:12:13
