淡色库蚊抗药性相关新基因—NYD-MLC2克

发布时间：2018-04-09 05:24

  本文选题：淡色库蚊　切入点：抗药性　出处：《南京医科大学》2006年硕士论文

【摘要】：【目的】 鉴定淡色库蚊溴氰菊酯抗性和敏感品系中肌球蛋白轻链2基因(NYD-MLC2)的表达差异，获取NYD-MCL2 cDNA全长序列，，并构建NYD-MLC2原核和昆虫表达载体，为进一步研究奠定基础。 【方法】 用荧光定量PCR法鉴定NYD-MLC2在淡色库蚊溴氰菊酯抗性和敏感品系中的表达差异。用RACE法从抗性品系中扩增NYD-MLC2 cDNA全长序列，对NYD-MLC2编码的蛋白进行生物信息学分析。用Blast软件在NCBI和SwissProt数据库中进行同源性比对，用Clustalw软件进行聚类分析，用EMBL pI／Mw tool计算等电点和分子量，Prot-Param分析NYD-MLC2基因编码的蛋白质的基本性质，Signal3.0软件进行信号肽分析，ProtScale软件进行疏水性分析，PSORT Ⅱ软件进行细胞定位分析，PatSearch软件进行基序分析，SMART软件进行蛋白质结构域分析，HNN软件分析二级结构，Tmpred软件进行跨膜区预测等。通过基因重组方法构建NYD-MLC2原核和昆虫表达载体。 【结果】 荧光定量PCR结果显示，NYD-MLC2在淡色库蚊溴氰菊酯抗性品系中的表达量是敏感品系的4.08倍。获得淡色库蚊NYD-MLC2 cDNA全长序列，其中完整开读框序列为630bp，经推导编码210个氨基酸(GenBank／NCBIDQ140391，2005)，与多个物种的肌球蛋白轻链2基因比对，同源性大于25％。成功构建NYD-MLC2原核和昆虫表达载体。 【结论】 NYD-MLC2在淡色库蚊溴氰菊酯抗性品系中高表达，提示NYD-MLC2与淡色库蚊溴氰菊酯抗性相关，值得进一步研究。
[Abstract]:[objective] to identify the difference of expression of myosin light chain 2 gene (NYD-MLC2) in Culex pipiens pallens deltamethrin resistance and sensitive strain, obtain the full-length NYD-MCL2 cDNA sequence, and construct NYD-MLC2 prokaryotic expression vector and insect expression vector, and lay a foundation for further study.[methods] the expression of NYD-MLC2 in deltamethrin resistant and sensitive strains of Culex pipiens pallens was identified by fluorescence quantitative PCR.The full-length sequence of NYD-MLC2 cDNA was amplified by RACE from resistant strains, and the protein encoded by NYD-MLC2 was analyzed by bioinformatics.Comparing the homology between NCBI and SwissProt database with Blast software, cluster analysis with Clustalw software and EMBL with EMBLPI/Mw tool calculation of isoelectric points and Molecular weight Prot-Param Analysis of the basic Properties of proteins encoded by NYD-MLC2 Gene; signal peptide analysis by signal peptide 3.0 software; hydrophobicity analysis by ProtScale software; cell localization analysis by PSORT 鈪

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