驴Zfy基因cDNA克隆及生物信息学分析

doi:10.16431/j.cnki.1671-7236.2018.01.005

摘要/Abstract

摘要：

本研究旨在对驴Zfy基因的cDNA序列进行克隆及生物信息学分析。根据GenBank中公布的家牛Zfy基因mRNA序列（登录号：NM_177491.1）设计特异性引物，运用RT-PCR技术获取驴Zfy基因的cDNA序列，并对Zfy基因CDS区核苷酸序列与蛋白质结构进行生物信息学分析。结果表明，驴Zfy基因CDS区序列长度为2 325 bp，编码774个氨基酸；蛋白质二级结构预测结果显示，Zfy蛋白没有信号肽及跨膜结构；驴Zfy基因CDS区序列与家牛、绵羊、人、家猫、家犬、马鹿、黑猩猩、藏羚羊的同源性分别为92.9%、92.6%、91.8%、93.8%、92.5%、92.3%、91.6%和92.6%；对驴和其他8种哺乳动物Zfy基因CDS区核苷酸序列构建的系统进化树显示，驴与绵羊、藏羚羊、家牛、马鹿亲缘关系较近，与家犬和家猫的亲缘关系稍远，与人和黑猩猩亲缘关系最远。本研究成功克隆出驴Zfy基因CDS区序列，为进一步研究驴Zfy基因的功能奠定了基础。

关键词: 驴; Zfy基因; 克隆; 生物信息学

Abstract:

This study was aimed to clone the cDNA sequence of Equus asinus Zfy gene and analyzed its genetic structure with bioinformatics. Specific primers were designed according to the mRNA sequence (accession No.:NM_177491.1) of Bos taurus Zfy gene published in GenBank, and Zfy gene were amplified and cloned by RT-PCR. The nucleotide sequence and protein structure of CDS region were analyzed through bioinformatic method. The results showed that the length of CDS region of Equus asinus Zfy gene was 2 325 bp, encoding 774 amino acids. Compared with Bos taurus, Ovis aries, Homo sapiens, Felis catus, Canis lupus, Cervus elaphus, Pan troglodytes and Pan tholops hodgosonii, the homology of the CDS region of Equus asinus Zfy gene were 92.9%, 92.6%, 91.8%, 93.8%, 92.5%, 92.3%, 91.6% and 92.6%, respectively. The phylogenetic tree, constructed from the CDS region of Equus asinus and eight other mammalian Zfy gene cDNA, displays that Equus asinus and Ovis aries, Pantholops hodgosonii, Bos taurus, Cervus elaphus had a closer genetic relationship, however the relationship was far away among Equus asinus, Felis catus and Canis lupus, and the relationship with Homo sapiens and Pan troglodytes was the farthest. In this study, we cloned the CDS region of Equus asinus Zfy gene, which laid the foundation for the further study of the function of Zfy gene.

Key words: Equus asinus; Zfy gene; cloning; bioinformatics

中图分类号:

营瑞文, 张永生, 夏欢, 王香祖, 李超程, 王绪海, 席继锋, 贾斌. 驴Zfy基因cDNA克隆及生物信息学分析[J]. 《中国畜牧兽医》, 2018, 45(1): 39-46.

YING Ruiwen, ZHANG Yongsheng, XIA Huan, WANG Xiangzu, LI Chaocheng, WANG Xuhai, XI Jifeng, JIA Bin. cDNA Cloning and Bioinformatics Analysis of Zfy Gene in Equus asinus[J]. , 2018, 45(1): 39-46.

参考文献

[1] PALMER C J, GALAN-CARIDAD J M, WEISBERG S P, et al. Zfx facilitates tumorigenesis caused by activation of the Hedgehog pathway[J]. Cancer Research, 2014, 74(20):5914-5924.
[2] SKINNER B M, SARGENT C A, CHURCHER C, et al. The pig X and Y chromosomes:Structure, sequence, and evolution[J]. Genome Research,2016,26(1):130-139.
[3] 刘帅兵. 绵羊Zfy基因干扰载体筛选及性控效果的验证[D]. 石河子:石河子大学,2014. LIU S B. The RNAi vector screening and verification of the effects of sexual control from Ovis arise Zfy gene[D]. Shihezi:Shihezi University, 2014.(in Chinese)
[4] 彭强, 班谦, 贾斌, 等. siRNA介导小鼠Zfy基因干扰的定量分析[J]. 石河子大学学报(自然科学版), 2011, 29(4):448-451. PENG Q,BAN Q,JIA B,et al. Quantitative analysis siRNA mediated silencing Zfy gene of Mus[J]. Journal of Shihezi University(Natural Science),2011,29(4):448-451.(in Chinese)
[5] 马世俊. 小鼠Zfx基因RNAi片段的筛选及性控效果的观察[D]. 石河子:石河子大学,2013. MA S J.The RNAi fragments screening and observation of the effects of sexual control from Mus Zfx gene[D]. Shihezi:Shihezi University,2013.(in Chinese)
[6] 纪俊明. 犬Zfx/Zfy基因干扰载体构建及性控效果的观察[D]. 石河子:石河子大学,2015. JI J M. The RNAi vecter construction and verification of the effects of sexual control from dog Zfx/Zfy gene[D]. Shihezi:Shihezi University,2015.(in Chinese)
[7] 李建华,罗军,张晓,等.奶山羊POU1F1基因CDS区的克隆、分析及原核表达[J]. 西北农林科技大学学报(自然科学版),2010,38(1):41-45. LI J H, LUO J, ZHANG X, et al. Cloning and analysis of POU1F1 gene in dairy goat and its prokaryotic expression[J]. Journal of Northwest A & F University (Natural Science Edition), 2010, 38(1):41-45. (in Chinese)
[8] 李元吉,杨明华,刘芳,等. 树鼩Tssk6基因cDNA克隆、组织表达谱及生物信息学分析[J].中国畜牧兽医,2017, 44(3):619-627. LI Y J,YANG M H,LIU F,et al. cDNA cloning,tissue expression profiles and bioinformatical analysis of Tssk6 gene in tree shrews[J]. China Animal Husbandry & Veterinary Medicine,2017, 44(3):619-627.(in Chinese)
[9] 李盛杰,杜晓华,罗玉柱,等.天祝白牦牛NGB基因的克隆及生物信息学分析[J].畜牧兽医学报,2013,44(3):395-398. LI S J, DU X H, LUO Y Z, et al. Cloning and bioinformatics analysis of NGB gene of Tianzhu White yak[J]. Chinese Journal of Animal and Veterinary Sciences, 2013, 44(3):395-398. (in Chinese)
[10] 刘虎岐,刘应保,宋云鹏,等. 淡色库蚊氯菊酯抗性相关基因PR-OP全长cDNA的克隆及生物信息学分析[J].西北农林科技大学学报(自然科学版), 2010,38(9):109-116. LIU H Q, LIU Y B, SONG Y P, et al. Cloning and bioinformatics analysis of full length cDNA of permethrin-resistance associated opsin gene PR-OP of Culex pipenspallens[J]. Journal of N & F University (Natural Science Edition), 2010, 38(9):109-116. (in Chinese)
[11] 孙雪婧,杜晓华,杨孝朴,等. 牦牛CYGB基因CDS区克隆与生物信息学分析[J]. 中国农业科学, 2014, 47(13):2690-2698. SUN X J, DU X H,YANG X P, et al. Cloning and bioinformatics analysis on CDS of CYGB gene in yak[J]. Scientia Agricultura Sinica, 2014,47(13):2690-2698. (in Chinese)
[12] ZHANG Y, XI J, JIA B, et al. RNAi as a tool to control the sex ratio of mouse offspring by interrupting Zfx/Zfy genes in the testis[J]. Mammalian Genome:Official Journal of the International Mammalian Genome Society, 2017, 28(3-4):100-105.
[13] VERNET N, MAHADEVAIAH S K, DE ROOIJ D G,et al. Zfy genes are required for efficient meiotic sex chromosome inactivation (MSCI) in spermatocytes[J]. Human Molecular Gengtics, 2016, 25(24):5300-5310.
[14] 张永生,席继峰,王香祖,等. Zfy基因与哺乳动物性别控制研究进展[J].畜牧与兽医,2016,48(12):105-108. ZHANG Y S,XI J F,WANG X Z,et al. Research progress of Zfy gene in mammals sex control[J].Animal Husbandry and Veterinary Medicine,2016,48(12):105-108.(in Chinese)
[15] ZWINGMAN T,ERICKSON R P,BOYER T,et al.Transcription of the sex-determining region genes Sry and Zfy in the mouse preimplantation embryo[J].Proceedings of the National Academy Science of the United States of America,1993,90(3):814-817.
[16] DECARPENTRIE F,VERNET N,MAHADEVAIAH S K,et al. Human and mouse Zfy genes produce a conserved testis-specific transcript encoding a zinc finger protein with a short acidic domain and modified transactivation potential[J]. Human Molecular Genetics,2012,21(12):2631-2645.
[17] VERNET N, MAHADEVAIAH S K, YAMAUCHI Y, et al. Mouse Y-linked Zfy1 and Zfy2 are expressed during the male-specific interphase between meiosis Ⅰ and meiosis Ⅱ and promote the 2nd meiotic division[J]. PLoS Genetics, 2014, 10(6):e1004444.
[18] NAKASUJI T, OGONUKI N, CHIBA T, et al. Complementary critical functions of Zfy1 and Zfy2 in mouse spermatogenesis and reproduction[J]. PLoS Genetics, 2017,13(1):e1006578.
[19] HAN S H,YANG B C,KO M S,et al. Length difference between equine Zfx and Zfy genes and its application for molecular sex determination[J]. Journal Assisted Reproduction Genetics,2010,27(12):725-728.
[20] ZHOU Q, BACHTROG D. Ancestral chromatin configuration constrains chromatin evolution on different iating sex chromosomes in drosophil[J]. PLoS Genetics, 2015, 11(6):e1005331.
[21] 魏丽敏,孙丽荣,张永生,等.马鹿Zfx/Zfy基因的克隆及序列分析[J].中国畜牧兽医,2016,43(9):2272-2278. WEI L M,SUN L R,ZHANG Y S,et al. Cloning and sequence analysis of Zfx/Zfy gene in Cervus elaphus[J].China Animal Husbandry & Veterinary Medicine,2016,43(9):2272-2278. (in Chinese)
[22] 贾浩,张小白,宋晓峰.人类胞内蛋白半衰期与其亚细胞定位的相关性研究[J]. 计算机与应用化学, 2011, 28(4):411-414. JIA H, ZHANG X B, SONG X F. Relationship between intracellular protein half-life and subcellular localiztion in human cells[J]. Computers and Applied Chemistry, 2011, 28(4):411-414.(in Chinese)
[23] 徐飞,成述儒,罗玉柱. 绵羊DRB1基因生物信息学分析[J].生物技术通报,2011,1:113-117. XU F, CHENG S R, LUO Y Z. Bioinformatics analysis of sheep DRB1 gene[J]. Biotechnology Bulletin, 2011, 1:113-117. (in Chinese)