牙鲆cbx2基因的分子特征与组织表达

doi:10.13560/j.cnki.biotech.bull.1985.2018-0995

生物技术通报 ›› 2019, Vol. 35 ›› Issue (4): 69-75.doi: 10.13560/j.cnki.biotech.bull.1985.2018-0995

牙鲆cbx2基因的分子特征与组织表达

王新艳¹, 张俊玲^1,2,3, 施志仪^1,2,3

1. 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;
2. 上海海洋大学水产科学国家级实验教学示范中心,上海 201306;
3. 上海海洋大学上海水产养殖工程技术研究中心,上海 201306

收稿日期:2018-11-19 出版日期:2019-04-26 发布日期:2019-05-05
作者简介:王新艳,女,硕士研究生,研究方向：生物化学与分子生物学;E-mail：18621092983@163.com
基金资助:
国家自然科学基金项目（41306128）

Molecular Characterization and Tissue Expression of cbx2 Gene in Paralichthys olivaceus

WANG Xin-yan¹, ZHANG Jun-ling^1,2,3, SHI Zhi-yi^1,2,3

1. Laboratory of Freshwater Aquatic Genetic Resources, Ministry of Agriculture,Shanghai Ocean University,Shanghai 201306;
2. National Demonstration Center for Experimental Fisheries Science Education,Shanghai Ocean University, Shanghai 201306;
3. Shanghai Engineering Research Center of Aquaculture,Shanghai Ocean University,Shanghai 201306

Received:2018-11-19 Published:2019-04-26 Online:2019-05-05

摘要/Abstract

摘要： 为鉴定牙鲆（Paralichthys olivaceus）色素框同源蛋白（Chromobox homolog,CBX）基因cbx2及探讨其在牙鲆性腺中的表达,通过PCR克隆和测序获得了cbx2 cDNA序列;利用多种生物信息学方法在线分析了牙鲆CBX2蛋白质的理化性质与空间结构,并比较了脊椎动物中cbx2的基因结构、氨基酸同源性、系统进化;运用RT-PCR技术分析了cbx2基因在牙鲆成体不同组织的表达情况。结果表明获得的牙鲆cbx2 cDNA序列包含1 584 bp的开放阅读框,编码527个氨基酸,其蛋白质分子量和等电点分别为56 578.98和10.03,是一种偏碱性的亲水性蛋白,该蛋白无信号肽,无跨膜区域,且编码该蛋白的氨基酸中以丝氨酸（Sr）的含量为最高;空间结构分析发现,牙鲆CBX2蛋白以α螺旋和无规则卷曲为主,其二级结构中含有1个染色质结构域和4个低复杂结构域,该染色质结构域常结合甲基化氨基酸残基。基因结构、氨基酸同源性和系统进化分析表明cbx2基因在脊椎动物进化中较为保守,牙鲆cbx2与尼罗罗非鱼（Oreochromis niloticus）、青鳉（Oryzias latipes）的基因结构更为相似,其氨基酸同源性与杜氏鰤（Seriola dumerili）最高,为89%。RT-PCR结果显示牙鲆cbx2基因在精巢的表达量最高,卵巢次之,而在其他组织中仅有较低的表达。这些结果提示了cbx2基因在牙鲆的性腺发育中可能具有重要作用。

关键词: 牙鲆, cbx2, 性腺, 生物信息学分析, RT-PCR

Abstract: In order to identify the cbx2 gene of chromobox homolog（CBX）and its expression in the gonad of Paralichthys olivaceus,the cDNA sequence of cbx2 gene was obtained by PCR cloning and sequencing. The physicochemical properties and spatial structure of CBX2 protein were analyzed online by bioinformatics methods,and the gene structure,amino acid homology and phylogenetic evolution of cbx2 in vertebrates were compared. The expression of cbx2 gene in different adult tissues of P. olivaceus was detected by RT-PCR. The results showed that the obtained cbx2 cDNA had an open reading frame of 1 584 bp,encoding 527 amino acids in P. olivaceus,and the molecular weight and isoelectric point of the reduced protein was 56 578.98 and 10.03,respectively. The CBX2 protein was an alkaline and hydrophilic protein,had no signal peptide and no transmembrane area,and the amino acid encoding the protein had the highest content of serine（Sr）. The spatial structure analysis revealed that the CBX2 protein was mainly composed of α-helix and random coil,and its secondary structure contained one chromatin domain and four low complexity regions,and the chromatin domain was often binding to methylated amino acids. The gene structure,amino acid homology,and phylogenetic analysis indicated that the cbx2 gene was more conserved in the evolution of vertebrate. The gene structure of cbx2 was more similar to that of Oreochromis niloticus and Oryzias latipes,and its amino acid homology was the highest with Seriola dumerili,which was 89%. The RT-PCR showed that the expression of cbx2 mRNA was the highest in the testis,the second one in the ovary,but quite low expressions in other tissues. All above results suggest that cbx2 gene plays a key role in the gonad development of P. olivaceus.

Key words: Paralichthys olivaceus, cbx2, gonad, bioinformatics analysis, RT-PCR

王新艳, 张俊玲, 施志仪. 牙鲆cbx2基因的分子特征与组织表达[J]. 生物技术通报, 2019, 35(4): 69-75.

WANG Xin-yan, ZHANG Jun-ling, SHI Zhi-yi. Molecular Characterization and Tissue Expression of cbx2 Gene in Paralichthys olivaceus[J]. Biotechnology Bulletin, 2019, 35(4): 69-75.

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牙鲆cbx2基因的分子特征与组织表达

Molecular Characterization and Tissue Expression of cbx2 Gene in Paralichthys olivaceus

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