猪CatSperB和CatSperG基因的克隆、表达及生物信息学

doi:10.3864/j.issn.0578-1752.2012.21.015

摘要/Abstract

摘要： 【目的】揭示猪CatSperB和CatSperG基因的存在、蛋白的结构特征、进化关系及时空表达特性。【方法】利用电子和分子克隆技术鉴定猪CatSperB和CatSperG基因全长cDNA，并利用定性RT-PCR和荧光定量RT-PCR进行CatSperB和CatSperG基因的时空表达研究。【结果】①分别获得了3 508 bp CatSperB和3 715 bp CatSperG电子转录子，分别包含3 330和3 483 bp开放阅读框，并经TA克隆测序验证，其CDS序列与人、牛、马和狗等的CatSperB和CatSperG基因的序列相似性在80%以上；②CatSperB分子质量为125.79 kD，为稳定蛋白；CatSperG分子质量为133.40 kD，为不稳定蛋白；③CatSperB和CatSperG 都包含 7 个通道蛋白保守的跨膜结构域，CatSperG蛋白C端含一个超螺旋结构，而CatSperB蛋白无明显的超螺旋结构信号；猪CatSperB和CatSperG与牛、狗和马的CatSperB和CatSperG蛋白同源关系较近，与人和小鼠的同源关系较远；④RT-PCR分析表明，CatSperB和CatSperG基因主要在睾丸中表达，但CatSperB在其它组织也有表达信号；⑤CatSperB和CatSperG基因mRNA表达水平在猪性发育的重要阶段，精子发生（60日龄）、初情期（90日龄）和性成熟（150日龄）前后都有显著提高（P＜0.05）。【结论】获得了猪CatSperB和CatSperG基因的cDNA克隆及其一系列生物信息学参数，揭示了CatSperB和CatSperG蛋白含7个保守的跨膜结构域及不同物种间的进化关系，证实CatSperB和CatSperG基因主要在睾丸表达，且其mRNA表达变化与公猪的性发育相一致。

关键词: 猪, CatSperB基因, CatSperG基因, 克隆, 时空表达

Abstract: 【Objective】The aim of the current study is to confirm the existence of porcine CatSperB and CatSperG genes, and investigate the protein structures, evolutionary relationship and the spatial-temporal expression profiles of CatSperB and CatSperG. 【Method】The in silico and molecular cloning was used to identify the full length cDNAs of porcine CatSperB and CatSperG, and the spatial-temporal expression profile was investigated by qualitative and fluorescence quantitative RT-PCR. 【Result】The in silico transcripts of 3 508 bp CatSperB and 3 715 bp CatSperG were identified, and they contain 3 330 bp and 3 483 bp ORFs of CatSperB and CatSperG, respectively, and the sequences were confirmed by TA cloning. The sequence similarity of coding sequences (CDS) of porcine CatSperB and CatSperG with human, cattle, horse, and dog, and other animals was above 80%. CatSperB is a 125.79 kD and stable protein, while CatSperG is a 133.40 kD and unstable protein. Both CatSperB and CatSperG contain seven conservative trans-membrane domains, and a coiled-coil motif was also identified in the C terminal of CatSperG, but this motif was not found in CatSperB. The porcine CatSperB and CatSperG displayed higher degree of homology with the orthologs of cattle, dog and horse, and lower homology with those of human and mouse. The RT-PCR analysis showed that CatSperB and CatSperG were detected mainly in testis, but CatSperB also expressed in other tissues. The mRNA expression of CatSperB and CatSperG was significantly improved at the around stage of spermatogenesis (Day 60), puberty (Day 90) and sex maturity (Day 150) (P＜0.05). 【Conclusion】The cDNA clones of CatSperB and CatSperG, and a series of bioinformatics parameters of these proteins were obtained. Seven conservative trans-membrane domains in CatSperB and CatSperG and the homology among species were revealed. Furthermore, results of this study had proved that CatSperB and CatSperG expressed mainly in tests, and the change of mRNA expression was paralleled with sexual development of boar.

Key words: pig, CatSperB gene, CatSperG gene, cloning, spatial-temporal expression

宋成义, 周家庆, 冯晓军, 谢雨琇, 李庆平, 吴晗, 高波, 王霄燕. 猪CatSperB和CatSperG基因的克隆、表达及生物信息学[J]. 中国农业科学, 2012, 45(21): 4465-4474.

SONG Cheng-Yi, ZHOU Jia-Qing, FENG Xiao-Jun, XIE Yu-Xiu, LI Qing-Ping, WU Han, GAO Bo, WANG Xiao-Yan. Cloning, Expression and Bioinformatics Analysis of Porcine CatSperB and CatSperG Genes[J]. Scientia Agricultura Sinica, 2012, 45(21): 4465-4474.

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