[1]庞春英,陆杏蓉,朱鹏,等.水牛透明带3基因启动子克隆及转录活性检测[J].江苏农业学报,2015,(06):1350-1356.[doi:doi:10.3969/j.issn.1000-4440.2015.06.024]
　PANG Chun-ying,LU Xing-rong,ZHU Peng,et al.Cloning and transcriptional activity of promoter of buffalo zone pellucida 3(ZP3) gene[J].,2015,(06):1350-1356.[doi:doi:10.3969/j.issn.1000-4440.2015.06.024]
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水牛透明带3基因启动子克隆及转录活性检测()
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江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2015年06期
页码:
1350-1356
栏目:
畜牧兽医·水产养殖
出版日期:
2015-12-31

文章信息/Info

Title:
Cloning and transcriptional activity of promoter of buffalo zone pellucida 3(ZP3) gene
作者:
庞春英; 陆杏蓉; 朱鹏; 邓廷贤; 段安琴; 陈明棠; 杨炳壮; 梁贤威
(中国农业科学院广西水牛研究所广西水牛遗传繁育重点实验室，广西南宁530001)
Author(s):
PANG Chun-ying; LU Xing-rong; ZHU Peng; DENG Ting-xian; DUAN An-qin; CHEN Ming-tang; YANG Bing-zhuang; LIANG Xian-wei
(Guangxi Buffalo Research Institute, Chinese Academy of Agricultural Science/Guangxi Key Laboratory of Buffalo Genetics, Breeding and Reproduction, Nanning 530001, China)
关键词:
水牛; ZP3基因; 克隆分析; 转录活性
Keywords:
buffalo; ZP3(zone pellucide 3) gene; cloning and bio-informatics analysis; transcriptional activity
分类号:
S823.8+3.2
DOI:
doi:10.3969/j.issn.1000-4440.2015.06.024
文献标志码:
A
摘要:
为了阐明水牛透明带3基因（ZP3）表达调控机理，通过PCR技术、载体构建和细胞转染等技术，对其转录活性进行了研究。结果表明：成功克隆得到广西本地水牛ZP3基因5′侧翼序列及部分编码区序列（CDS）区序列，共3 081 bp；广西本地水牛ZP3核苷酸序列与河流型水牛、黄牛、绵羊和山羊的相似性分别为99%、96％、92％和92%；在ZP3翻译起始位点上游-147 bp至-196 bp处存在TATA box，启动子区存在GATAs、Foxo3、Nobox、Stat3、Stat4、Stat5a、Stat5b、Stat6和YY1等反式作用因子结合位点，其中Stats家族在ZP3启动子区存在多个结合位点，且同一位点又存在多个Stats结合的情况；水牛ZP3启动子不能启动绿色荧光蛋白(Enhanced green fluorescent protein, EGFP)在HEK-293T细胞中的表达，但能启动其在CHO细胞中的表达，且启动子活性比CMV启动子弱。
Abstract:
To identify the expression mechanism of buffalo zone pellucida 3 gene (〖WTBX〗ZP3), PCR, vector construction and cell transfection were applied to clone the gene and study its transcriptional activity. 3 081 bp ZP3 gene 5′ flanking and partial coding region sequence (CDS) of Guangxi local swamp buffalo was successfully cloned and sequenced. The nucleotide sequence of 〖WTBX〗ZP3 gene shared 99%, 96%, 92% and 92% similarities with those of river type buffalo, cattle, sheep and goat, respectively. There existed a TATA box in the location of -147 bp to -196 bp from the upstream of the translation initiation site and trans-acting factor binding sites for GATAs, Foxo3, Nobox, Stat3, Stat4, Stat5a, Stat5b, Stat6 and YY1. Multiple binding sites were presented in the promoter region of 〖WTBX〗ZP3 gene for Stats family genes, and multiple Stats were binding together in the same site of 〖WTBX〗ZP3 promoter. The promoter of 〖WTBX〗ZP3 gene could initiate EGFP expression in CHO cell lines rather than in HEK-293T cells, with weaker activity than CMV promoter in CHO cell lines. 

参考文献/References:

[1]NANDI S, RAGHU H M, RAVINDRANATHA B M, et al. Production of buffalo (Bubalus bubalis) embryos in vitro: premises and promises[J]. Reprod Domest Anim, 2002, 37(2): 65-74.
[2]ZICARELLI L, ESPOSITO L, CAMPANILE G, et al. Effects of using vasectomized bulls in artificial insemination practice on the reproductive efficiency of Italian buffalo cows[J]. Anim Reprod Sci, 1997,47(3): 171-180.
[3]MANJUNATHA B M, RAVINDRA J P, GUPTA P S, et al. Oocyte recovery by ovum pick up and embryo production in river buffaloes (Bubalus bubalis)[J]. Reprod Domest Anim, 2008, 43(4): 477-480.
[4]DI FRANCESCO S, NOVOA M V, VECCHIO D, et al. Ovum pick-up and in vitro embryo production (OPU-IVEP) in Mediterranean Italian buffalo performed in different seasons[J]. Theriogenology, 2012,77(1): 148-154.
[5]PONTES J H, MELO STERZA F A, BASSO A C, et al. Ovum pick up, in vitro embryo production, and pregnancy rates from a large-scale commercial program using Nelore cattle (Bos indicus) donors[J]. Theriogenology, 2011, 75(9): 1640-1646.
[6]崔奎青,刘庆友,李秀林,等. 基于线粒体控制区变异的水牛群体遗传分析[J]. 华南农业大学学报, 2013, 34(1): 76-82.
[7]崔奎青,张会娜,刘帅,等. 沼泽型水牛NANOG基因5'调控序列克隆与功能分析[J]. 畜牧兽医学报, 2013, 44(3): 387-394.
[8]邓彦飞,刘真真,李云芳,等. 不同逆转录载体系统应用于水牛胎儿成纤维细胞转基因的探索[J]. 华南农业大学学报, 2013, 34(4): 558-563.
[9]龚云,乔树叶,林浪,等. 水牛〖WTBX〗YY1基因克隆及其shRNA片段的筛选[J]. 畜牧兽医学报, 2014, 45(1): 62-68.
[10]苏节,刘庆友,朱鹏,等. 沼泽型水牛〖WTBX〗CYP19A1基因克隆、序列分析及组织表达研究[J]. 畜牧兽医学报, 2013, 44(4): 514-521.
[11]SHI J W, SHENG J Q, PENG K, et al. Expression pattern of the zona pellucida 3 (〖WTBX〗ZP3) gene during ovarian development and the location of ZP3 protein in oocytes in a natural, wild triploid crucian carp mutant, Carassius auratus var. Pingxiangnensis[J]. Genet Mol Res, 2013, 12(4): 5640-5650.
[12]GUPTA S K, BHANDARI B, SHRESTHA A, et al. Mammalian zona pellucida glycoproteins: structure and function during fertilization[J]. Cell Tissue Res, 2012, 349(3): 665-678.
[13]YONEZAWA N, KANAI-KITAYAMA S, KITAYAMA T, et al. Porcine zona pellucida glycoprotein ZP4 is responsible for the sperm-binding activity of the ZP3/ZP4 complex[J]. Zygote, 2012, 20(4): 389-397.
[14]LAN Z J, XU X, COONEY A J. Differential oocyte-specific expression of Cre recombinase activity in GDF-9-iCre, Zp3cre, and Msx2Cre transgenic mice[J]. Biol Reprod, 2004, 71(5): 1469-1474.
[15]DE VRIES W N, BINNS L T, FANCHER K S, et al. Expression of Cre recombinase in mouse oocytes: a means to study maternal effect genes[J]. Genesis, 2000, 26(2): 110-112.
[16]LIANG L, SOYAL S M, DEAN J. FIGalpha, a germ cell specific transcription factor involved in the coordinate expression of the zona pellucida genes[J]. Development, 1997, 124(24): 4939-4947.
[17]EPIFANO O, LIANG L F, FAMILARI M, et al. Coordinate expression of the three zona pellucida genes during mouse oogenesis[J]. Development, 1995, 121(7): 1947-1956.
[18]LIU C, LITSCHER E S, MORTILLO S, et al. Targeted disruption of the mZP3 gene results in production of eggs lacking a zona pellucida and infertility in female mice[J]. Proc Natl Acad Sci USA, 1996, 93(11): 5431-5436.
[19]EPIFANO O, LIANG L F, DEAN J. Mouse Zp1 encodes a zona pellucida protein homologous to egg envelope proteins in mammals and fish[J]. J Biol Chem, 1995, 270(45): 27254-27258.
[20]SHI J, PENG K, SHENG J, et al. A specific genomic organization and a novel promoter sequence for both 〖WTBX〗ZP2 and 〖WTBX〗ZP3 gene expressions in the Pingxiang red transparent crucian carp, Carassius auratus var. pingxiangnensis[J]. Comp Biochem Physiol Part D Genomics Proteomics, 2013, 8(4): 275-282.
[21]LEWANDOSKI M, WASSARMAN K M, MARTIN G R. Zp3-cre, a transgenic mouse line for the activation or inactivation of loxP-flanked target genes specifically in the female germ line[J]. Curr Biol, 1997, 7(2): 148-151.

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备注/Memo

备注/Memo:
收稿日期：2015-03-24 基金项目：农业部转基因重点项目（2014ZX08010-012B）；国际先进技术引进与合作研究开发项目（14123001-5） 作者简介：庞春英（1972-），女，广西博白人，本科，助理研究员，研究方向为水牛遗传育种与胚胎生物学技术。(Tel)15277012910；(E-mail)pangcy800@163.com 通讯作者：梁贤威，（E-mail）liangbri@126.com
更新日期/Last Update: 2015-12-31