中华绒螯蟹几丁质酶基因HXchit全长cDNA克隆及其在蜕皮过程中的表达分析
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上海海洋大学 农业部淡水水产种质资源重点实验室, 上海 201306

作者简介:

姚琴琴((1988.11–)), 女, 硕士研究生, 主要从事中华绒螯蟹分子营养学研究. E-mail: a1017604175@163.com

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S972.1Q959; S96

基金项目:

国家863高技术研究发展计划资助项目(2012AA10A409-5); ; 国家自然科学基金资助项目(31472287; 31272677); 国家农业科技成果转化项目(2012GB2C000147) ;; 上海教委知识服务平台项目(EF1206); 国家自然科学基金项目(31272677); 上海市科委科技合作专项((13340721500)); ; 上海市科委优秀学术带头人项目((12XD1402700))。.


Full length cDNA cloning of the chitinase gene (HXchit) and analysis of expression during the molting cycle of the Chinese mitten crab, Eriocheir
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Key Laboratory of Freshwater Aquatic Genetic Resources, Ministry of Agriculture; Shanghai Ocean University, Shanghai 201306, China

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    摘要:

    (在中华绒螯蟹的同一蜕皮阶段不同组织、不同蜕皮阶段主要几个组织的表达情况。结果显示(ORF) 1 470 bp, , 和相似性为表皮中期、AB肌肉中期最高。结果表明

    Abstract:

    undergoes several molts in its lifetime. We cloned the chitinase gene from using reverse transcriptase polymerase chain reaction and rapid-amplification of cDNA ends. Primers were designed based on the conserved sequence of chitinase from was 2042 bp, including a 1470 bp ORF (encoded 490 amino acid residues), a 35 bp 5′-UTR, and a 537 bp 3′-UTR. The calculated molecular weight and isoelectric point were 53.44 kD and 4.41, respectively. A homology analysis using BLASTn and BLASTx revealed that the S. serrata. The phylogenetic tree of the chitinase gene generated by the Neighbor Joining method suggested that . The expression of was analyzed by real-time fluorescent quantitative PCR. -mRNA was expressed in all tissues examined and levels were highest in the hepatopancreas, with only low levels of expression in the muscle, stomach, gill, and cuticle, and trace levels of expression in the heart, intestine, thoracic ganglion, and brain ganglion. -mRNA expression in the hepatopancreas peaked during the D stage, and there was a significant difference (<0.05) in expression in the hepatopancreas between stages E, AB, and C. In the cuticle, the level of -mRNA expression increased gradually during the molting cycle, and was significantly higher during D stage than during all other stages (<0.05). Expression levels were relatively low in the stomach throughout the molt cycle. In muscle, -mRNA expression was highest during C stage, but there was no difference in the level of expression between the different molting stages ( is a form of acid chitinase and is expressed abundantly in the hepatopancreas. may play a role in the digestion of chitinous food and the degradation of endogenous chitin during the molting cycle of . Our results lay a foundation for research into the biological function and regulation of chitinase in .

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姚琴琴,杨志刚,王瑶,郭子好,刘启彬,施秋燕,成永旭.中华绒螯蟹几丁质酶基因HXchit全长cDNA克隆及其在蜕皮过程中的表达分析[J].中国水产科学,2015,22(2):185-195
YAO Qinqin, YANG Zhigang, WANG Yao, GUO Zihao, LIU Qibin, SHI Qiuyan, CHENG Yongxu. Full length cDNA cloning of the chitinase gene (HXchit) and analysis of expression during the molting cycle of the Chinese mitten crab, Eriocheir[J]. Journal of Fishery Sciences of China,2015,22(2):185-195

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  • 在线发布日期: 2015-06-29
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