三疣梭子蟹Δ6脂肪酸去饱和酶基因全长cDNA克隆与组织表达

doi:10.13560/j.cnki.biotech.bull.1985.2015.09.019

摘要/Abstract

摘要： 旨在探讨三疣梭子蟹高度不饱和脂肪酸自身合成能力，探究三疣梭子蟹HUFA生物合成途径。采用cDNA末端快速扩增(RACE)技术克隆得到三疣梭子蟹△6去饱和酶cDNA 全长序列，并利用荧光定量PCR技术进行肝胰腺、肠道、鳃等8种组织的表达分析。通过分析序列表明，基因序列全长2 875 bp，其中5'非编码区长465 bp，3'非编码区长1 078 bp，开放阅读框(ORF)长1 332 bp，编码443个氨基酸；并且编码的蛋白序列具有典型的去饱和酶特性：3个组氨酸保守区，一个N端细胞色素b5结构域以及一个血红素结合的HPGG结构域。荧光定量PCR结果显示，Δ6脂肪酸去饱和酶基因在三疣梭子蟹多个组织中均有表达，在肝胰腺中表达量最高，其次是肠道和肌肉，心脏中表达最少。结果表明三疣梭子蟹具有△6去饱和酶。

关键词: 三疣梭子蟹, 脂肪酸去饱和酶, 组织表达分析

Abstract: The goal of this work is to investigate the self-synthesis capability of highly unsaturated fatty acid(HUFA)and understand the synthesis pathway of HUFA in Portunus trituberculatus. Full-length cDNA of gene for Δ6-desaturase fatty acyl in P. trituberculatus was cloned by rapid amplification of cDNA ends(RACE), and the expressions in 8 tissues such as hepatopancreas, intestine and gill etc. were analyzed by real-time quantitative PCR(qRT-PCR). Analysis of the full-length cDNA of gene for Δ6-desaturase fatty acyl revealed that it was 2 875 bp, including 465 bp of 5' UTR and 1 078 bp of 3' UTR, and encoded 443 amino acids. The presumed protein sequence had a typical desaturase structure：3 histidine-rich motifs, a b5 domain of an N terminal cytochrome and a heme-binding HPGG domain. qRT-PCR showed that gene for Δ6-desaturase fatty acyl expressed in all 8 tissues while the highest in hepatopancreas, less in intestine and muscle, and the least in the heart. The results indicate that P. trituberculatus possesses Δ6-desaturase fatty acyl.

Key words: Portunus trituberculatus, fatty acid desaturase, tissue expression analysis

施秋燕, 杨志刚, 王伟, 姚琴琴, 王瑶, 成永旭. 三疣梭子蟹Δ6脂肪酸去饱和酶基因全长cDNA克隆与组织表达[J]. 生物技术通报, 2015, 31(9): 138-145.

Shi Qiuyan, Yang Zhigang, Wang Wei, Yao Qinqin, Wang Yao, Cheng Yongxu. Cloning and Tissue Expression of Full-length cDNA in Gene Encoding Δ6-desaturase Fatty Acyl of Portunus trituberculatus[J]. Biotechnology Bulletin, 2015, 31(9): 138-145.

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