口虾蛄高血糖激素基因全长cDNA的克隆及表达分析

刘海映; 张秀芹; 隋宥珍; 李宏俊; 邢坤; 姜玉声; 杨贵福; 陈雷; 张娜

doi:10.3969/j.issn.0253-4193.2016.06.011

口虾蛄高血糖激素基因全长cDNA的克隆及表达分析

doi: 10.3969/j.issn.0253-4193.2016.06.011

1.
大连海洋大学辽宁省海洋生物资源恢复与生境修复重点实验室, 辽宁大连 116023
2.
国家海洋环境监测中心, 辽宁大连 116023
3.
大连海洋大学农业部北方海水增养殖重点实验室, 辽宁大连 116023

基金项目: 国家海洋局海洋公益项目(200905019-4); 国家自然科学基金资助项目(41006079)。

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出版历程
- 收稿日期: 2015-07-01

Molecular cloning and expression of a full length cDNA of encoding crustacean hyperglycemic hormone(CHH)in Oratosquilla oratoria

1.
Key Laboratory of Marine Bio-resources Restoration and Habitat Reparation in Liaoning Province, Dalian Ocean University, Dalian 116023, China
2.
National Marine Environmental Monitoring Center, Dalian 116023, China
3.
Key Laboratory of Mariculture & Stock Enhancement in North China's Sea, Ministry of Agriculture, Dalian Ocean University, Dalian 116023, China

摘要

摘要: 本文首次采用RACE技术获得口虾蛄(Oratosquilla oratoria)眼柄部高血糖激素(crustacean hyperglycemic hormone,CHH)基因cDNA全长序列,共2 421 bp,5'UTR长度为180 bp,3'UTR 为1 821 bp,开放阅读框(ORF)长420 bp,编码139个氨基酸。预测蛋白二级结构α-螺旋23.74%,延伸链占23.02%,无规则卷曲占53.24%,预测分子量为15.47 kD,等电点(pI)为7.049。CHH成熟肽包含6个CHH家族中位置保守的Cys残基,C端为GK,推断为ES型CHH基因。聚类分析表明:口虾蛄CHH和十足目CHH聚为同一个分支的两亚群,具有较近的亲缘关系。荧光定量PCR分析结果表明,CHH基因在肠中表达量最高,在眼柄、触角中有较高的表达量,在肌肉中表达量相对较低,在卵巢中表达最低,而在精巢中不表达。
- 口虾蛄 /
- 高血糖激素CHH /
- 分子克隆 /
- 荧光定量PCR /
- 表达分析
Abstract: Total RNA in eyestalk of Oratosquilla oratoria was extracted and cDNA sequence of crustacean hyperglycemic hormone (CHH) were obtained by RACE for the first time. The result showed that the full length of the CHH gene is 2 421 bp,containing an 180 bp 5'-untranslated region (UTR),and 1 821 bp 3'UTR,and 420 bp open reading flame (ORF) which encoded 139 amino acids. Prediction of protein structure was composed of 23.74% α-helix,23.02% extension chain,53.24% random coil,and its molecular mass is 15 470 Daltons with an estimated pI of 7.049.The mature peptide contained six conserved Cys residues in CHH family. The C-terminus mature peptide was GK which has the same characteristics with ES-CHH. Based on protein similarity comparison,the separated CHH gene was classified into the type I CHH family of peptides. Phylogenetic analysis showed that CHH of O. oratoria and CHH of decapod species formed two subgroups on the same branch of the tree. The relative expression of CHH gene in different organizations was tested by real time fluorescent quantitative PCR,the expression was highest in the intestine,and then the eyestalk,antenna and brain,the expression is negligible in muscle and ovary,and no expression in testis.
- Oratosquilla oratoria /
- crustacean hyperglycemic hormone /
- molecular cloning /
- RT PCR /
- expression analysis

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