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利用生物信息学方法鉴定致倦库蚊23条几丁质酶基因,并对它们进行命名、分组,以及基因结构、蛋白结构域、蛋白理化特性和系统发育分析,随后使用q PCR方法检测Cq Cht10在致倦库蚊生长各时期的相对表达量。结果表明,致倦库蚊几丁质酶基因长度为1 360~19 218 bp,蛋白长度为3532 681 aa,分子量为39.557~279.503 k D,等电点为4.768.69。它们属于GH18家族,每条几丁质酶包含信号肽区域、催化结构域、几丁质结合域和连接区域,但包含的数量不同。此外,每条几丁质酶的催化结构域均包含保守序列FDGXDLDWEYP。本研究将致倦库蚊几丁质酶划分为12组,组Ⅰ~组Ⅶ沿用早期研究的分组标准,此外将组Ⅷ更名为组Ⅸ,再新增4个分组。Cq Cht10的q PCR实验结果显示,该基因在Ⅱ龄~Ⅳ龄幼虫期呈高表达,在蛹期和成虫期呈低表达,而在卵期和Ⅰ龄幼虫期呈微量表达,此外在雌蚊和雄蚊中无显著差异表达,提示该基因与致倦库蚊的蜕皮相关,且与性别关联不大。本研究的分析结果能够为致倦库蚊几丁质酶基因的功能研究提供有价值的信息。
Abstract：There were 23 chitinase genes identified in Culex quinquefasciatus via bioinformatics methods, of which the gene structures, protein domains, protein physicochemical characteristics and develop system were analyzed, following with gene nominating and classification. Then the relative expression of Cq Cht10 in different developmental stages of C. quinquefasciatus was determined by q PCR. The results showed that Chitinase of C.quinquefasciatus was with gene in length of 1 360~19 218 bp, proteins in length of 3532 681 amino acid, molecular weight of 39.557~279.503 k D, and the p I of 4.768.69. They were belonged to GH18 gene family, and each was composed by different numbers of signal peptide, catalytic domain, chitin binding domain and linker region.In addition, a conserved region FDGXDLDWEYP located in the catalytic domain among all the chitinase proteins.We assigned these genes into 12 groups, in which 7 groups(GroupⅠ~Ⅶ) maintained base on early reference, 1 group altered the name from Group Ⅷ to Group Ⅸ, and 4 new groups added. q PCR results displayed the gene Cq Cht10 highly expressed in Ⅱ~Ⅳ instars larva, lowly expressed in pupae and adult, and barely expressed in eggs and Ⅰinstars larva, and had no significant difference between female and male. These provided insights into the relationship with molting and the irrelative with their gender. Our study would provide valuable information for the functional research of chitinase proteins in C. quinquefasciatus.
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基本信息:
DOI：10.13417/j.gab.035.000797
中图分类号:R384.1
引用信息:
[1]赵文静,张晶,张春林,等.致倦库蚊几丁质酶家族基因的鉴定与分析[J].基因组学与应用生物学,2016,35(04):797-805.DOI:10.13417/j.gab.035.000797.
基金信息:
贵州省科技厅黔科合LH字[2015]7363号;; 贵州医科大学院基金合同字2013-06基金共同资助
2016-04-25
2016-04-25