Scientia Agricultura Sinica ›› 2012, Vol. 45 ›› Issue (6): 1183-1190.doi: 10.3864/j.issn.0578-1752.2012.06.017

• ANIMAL SCIENCE·RESOURCE INSECT • Previous Articles     Next Articles

Cloning of Dairy Goat Prm1 Gene CDS and Construction of the Eukaryotic Expression Vector

 LI  Ming-Zhao, LIU  Chao, SUN  Jun-Wei, ZHU  Hai-Jing, CAO  Hui, LU Xiao , CHOU  Pu-Bin, HUA  Jin-Lian   

  1. 西北农林科技大学动物医学院/陕西省干细胞工程技术研究中心/农业部动物生物技术重点开放性实验室,陕西杨凌 712100
  • Received:2011-05-09 Online:2012-03-15 Published:2011-08-18

Abstract: 【objective】The objective of the experiment is to study the Prm1 gene of dairy goat, and for further research on dairy goat spermatogenesis.【Method】The CDS of Prm1 gene was cloned by PCR from Guanzhong dairy goat testis tissue. The obtained sequence was constructed into the eukaryotic expression vector pIRES-GFP, and then transfected to GC1 cell line and human umbilical cord cells. PCR and immunohistochemistry were used to test the expression of Prm1 at different ages of dairy goat testis. 【Result】In dairy goats, the expression of Prm1 gene was high and specific in adult testis, located only in the head of sperm. The CDS of Prm1 gene of Guanzhong dairy goat was amplified and compared with the sequences of other species. The results showed that they shared a high homology, being 97.4% between dairy goats and bovine. The Prm1 gene was overexpressed in GC1 cells and human umbilical cord cells to confirm its function. 【Conclusion】 The complete CDS of dairy goat Prm1 gene and its expression patterns were obtained.

Key words: Guanzhong dairy goat, Prm1 gene, gene cloning, expression pattern

[1]Dahm R. Friedrich Miescher and the discovery of DNA. Developmental Biology, 2005, 276: 274-266.

[2]费仁仁, 邓锦松, 纪  林, 刘  平, 朱  斌,  陈  晖, 阎月敏, 王莎丽, 陈啸梅, 章静波, 薛社普. 大鼠精子细胞变态期核蛋白转换的研究. 解剖学报, 1997, 28(4): 388-392.

Fei R R, Deng J S, Ji L, Liu P, Zhu B, Chen H, Yan Y M, Wang S L, Chen X M, Zhang J B, Xue S P. Studies on rat spermatid nuclear protein transition. Acta Anatomica Sinica, 1997, 28(4): 388-392. (in Chinese)

[3]Klemm U, Lee C H, Burfeind P, Hake S, Engel W. Nucleotide sequence of a cDNA ecoding rat protamine and the haploid expression of the gene during rat spermatogenesis. Biological Chemistry Hoppe-Seyler, 1989, 370: 293-301.

[4]Sassone-Corsi P. Unique chromatin remodeling and transcriptional regulation in spermatogenesis. Science, 2002, 296(5576): 2176-2176.

[5]Steger K, Pauls K, Klonisch T, Franke F E, Bergmann M. Expression of protamine-1 and -2 mRNA during human spermiogenesis. Molecular Human Reproduction, 2000, 6(3): 219-225.

[6]Ramm S A, Oliver P L, Ponting C P, Stockley P, Emes R D. Sexual selection and the adaptive evolution of mammalian ejaculate proteins. Molecular Biology and Evolution, 2008, 25(1): 207-219.

[7]Tanaka H, Miyagawa Y, Tsujimura A, Matsumiya K, Okuyama A, Nishimune Y. Single nucleotide polymorphisms in the protamine-1 and -2 genes of fertile and infertile human male populations. Molecular Human Reproduction, 2003, 9(2): 69-73.

[8]Imken L, Rouba H, EI Houate B, Louanjli N, Barakat A, Chafik A, McElreavey K. Mutations in the protamine locus: association with spermatogenic failure? Molecular Human Reproduction, 2009, 15(11): 733-738.

[9]Ravel C, Chantot-Bastaraud S, El Houate B, Berthaut I, Verstraete L, de Larouziere V, Lourenço D, Dumaine A, Antoine J M. Mutations in the protamine1 gene associated with male in fertility. Molecular Human Reproduction, 2007, 13(7): 461-464.

[10]Steger K, Fink L, Failing K, Bohle R M, Kliesch S, Weidner W, Bergmann M. Decreased protamine-1 transcript levels in testes from infertile men. Molecular Human Reproduction, 2003, 9(6): 331-336.

[11]Iguchi N, Yang S, Lamb D J. An SNP in protamine 1: a possible genetic cause of male infertility? Medical Genetics, 2006, 43: 362-364.

[12]Wykes S M, Krawetz S A. The structural organization of sperm chromatin. Biological Chemistry, 2003, 278(32): 29471-29477.

[13]Zhao M, Shirley C R, Mounsey S, Meistrich M L. Nucleoprotein transitions during spermiogenesis in mice with transition nuclear protein Tnp1 and Tnp2 mutations. Biology of Reproduction, 2004, 71: 1016-1025.

[14]Mitchell V, Steger K, Marchetti C, Herbaut J C, Devos P, Rigot J M. Cellular expression of protamine1 and 2 transcripts in testicular spermatids from azoospermic men submitted to TESE–ICSI. Molecular Human Reproduction, 2005, 11(5): 373-379.

[15]Lee K, Haugen H S, Clegg C H, Braun R E. Premature translation of protamine 1 mRNA causes precocious nuclear condensation and arrests spermatid differentiation in mice. Proceedings of the National Academy of Sciences of the United States of America, 1995, 92: 12451-12455.

[16]蒙学莲, 郑亚东, 才学鹏. 鱼精蛋白的研究现状及应用. 黑龙江畜牧兽医, 2007, 5: 26-27.

Meng X L, Zheng Y D, Cai X. The present condition and application of protamine. Heilongjiang Animal Science and Veterinary Medicine, 2007, 5: 26-27. (in Chinese)

[17]Bower P A, Yelick P C, Hecht N B. Both P1 and P2 protaminegenes are expressed in mouse, hamster, and rat. Biology of Reproduction, 1987, 37: 479-488.

[18]Reeves R H, Gearhart J D, Hecht N B, Yelick P, Johnson P, O’Brien S J. Mapping of PRM 1 to human chromosome 16 and tight linkage of prm-1 and prm-2 on mouse Chromosome 16. Heredity, 1989, 80: 442-446.

[19]Threadgill D S, Kraus J P, Krawetz S A, Womack J E. Evidence for the evolutionary origin of human chromosome 21 from comparative gene mapping in the cow and mouse. Proceedings of the National Academy of Sciences of the United States of America, 1991, 88: 154-158.

[20]Casas-Carrillo E, Prill-Adams A, Price S G, Clutter A C, Kirkpatrick B W. Mapping genomic regions associated with growth rate in pigs. Animal Science, 1997, 75: 2047-2053.

[21]Gilbert I, Bissonnette N, Boissonneault G, Vallée M, Robert C. A molecular analysis of the population of mRNA in bovine spermatozoa. Reproduction, 2007, 133: 1073-1086.

[22]陈  宇, 费仁仁, 吴小芳, 葛晔华, 陈  晖, 马  静, 韩代书, 薛社普. 大鼠精蛋白基因在 MEL细胞中的表达. 中国医学科学院学报, 2002, 24(6): 601-605.

Chen Y, Fei R R, Wu X F, Ge Y H. Chen H, Ma J, Han D S, Xue S. Expression of rat protamine gene in MEL cells. Acta Academiae Medicinae Sinicae, 2002, 24(6): 601-605. (in Chinese)

[23]Tanaka H, Baba T. Gene expression in spermatogenesis. Cellular and Molecular Life Sciences, 2005, 62: 344-354.

[24]Balhorn R. The protamine family of sperm nuclear proteins. Genome Biology, 2007, 8(9): 227-227.
[1] GU LiDan,LIU Yang,LI FangXiang,CHENG WeiNing. Cloning of Small Heat Shock Protein Gene Hsp21.9 in Sitodiplosis mosellana and Its Expression Characteristics During Diapause and Under Temperature Stresses [J]. Scientia Agricultura Sinica, 2023, 56(1): 79-89.
[2] LI YuZe,ZHU JiaWei,LIN Wei,LAN MoYing,XIA LiMing,ZHANG YiLi,LUO Cong,HUANG Gui Xiang,HE XinHua. Cloning and Interaction Protein Screening of RHF2A Gene from Xiangshui Lemon [J]. Scientia Agricultura Sinica, 2022, 55(24): 4912-4926.
[3] ZHANG YunXiu,JIANG Xu,WEI ChunXue,JIANG XueQian,LU DongYu,LONG RuiCai,YANG QingChuan,WANG Zhen,KANG JunMei. The Functional Analysis of High Mobility Group MsHMG-Y Involved in Flowering Regulation in Medicago sativa L. [J]. Scientia Agricultura Sinica, 2022, 55(16): 3082-3092.
[4] QU Cheng,WANG Ran,LI FengQi,LUO Chen. Cloning and Expression Profiling of Gustatory Receptor Genes BtabGR1 and BtabGR2 in Bemisia tabaci [J]. Scientia Agricultura Sinica, 2022, 55(13): 2552-2561.
[5] ZHANG Li,ZHANG Nan,JIANG HuQiang,WU Fan,LI HongLiang. Molecular Cloning and Expression Pattern Analysis of NPC2 Gene Family of Apis cerana cerana [J]. Scientia Agricultura Sinica, 2022, 55(12): 2461-2471.
[6] ZHANG Lu,ZONG YaQi,XU WeiHua,HAN Lei,SUN ZhenYu,CHEN ZhaoHui,CHEN SongLi,ZHANG Kai,CHENG JieShan,TANG MeiLing,ZHANG HongXia,SONG ZhiZhong. Identification, Cloning, and Expression Characteristics Analysis of Fe-S Cluster Assembly Genes in Grape [J]. Scientia Agricultura Sinica, 2021, 54(23): 5068-5082.
[7] TAN YongAn,JIANG YiPing,ZHAO Jing,XIAO LiuBin. Expression Profile of G Protein-Coupled Receptor Kinase 2 Gene (AlGRK2) and Its Function in the Development of Apolygus lucorum [J]. Scientia Agricultura Sinica, 2021, 54(22): 4813-4825.
[8] WANG Na,ZHAO ZiBo,GAO Qiong,HE ShouPu,MA ChenHui,PENG Zhen,DU XiongMing. Cloning and Functional Analysis of Salt Stress Response Gene GhPEAMT1 in Upland Cotton [J]. Scientia Agricultura Sinica, 2021, 54(2): 248-260.
[9] XU HuanHuan,LI Yi,GAO Wei,WANG YongQin,LIU LeCheng. Cloning and Identification of γ-Glutamyl Transpeptidase AcGGT Gene from Onion (Allium cepa) [J]. Scientia Agricultura Sinica, 2021, 54(19): 4169-4178.
[10] SHI GuoLiang,WU Qiang,YANG NianWan,HUANG Cong,LIU WanXue,QIAN WanQiang,WAN FangHao. Gene Cloning, Expression Pattern and Molecular Characterization of Chitin Deacetylase 2 in Cydia pomonella [J]. Scientia Agricultura Sinica, 2021, 54(10): 2105-2117.
[11] TAN YongAn,ZHAO XuDong,JIANG YiPing,ZHAO Jing,XIAO LiuBin,HAO DeJun. Cloning, Preparation of Antibody and Response Induced by 20-Hydroxyecdysone of Target of Rapamycin in Apolygus lucorum [J]. Scientia Agricultura Sinica, 2021, 54(10): 2118-2131.
[12] KunNeng ZHOU,JiaFa XIA,Peng YUN,YuanLei WANG,TingChen MA,CaiJuan ZHANG,ZeFu LI. Transcriptome Research of Erect and Short Panicle Mutant esp in Rice [J]. Scientia Agricultura Sinica, 2020, 53(6): 1081-1094.
[13] XING QiKai,LI LingXian,CAO Yang,ZHANG Wei,PENG JunBo,YAN JiYe,LI XingHong. Prediction and Analysis of Candidate Secreted Proteins from the Genome of Lasiodiplodia theobromae [J]. Scientia Agricultura Sinica, 2020, 53(24): 5027-5038.
[14] HAO ShuLin,CHEN HongWei,LIAO FangLi,LI Li,LIU ChangYan,LIU LiangJun,WAN ZhengHuang,SHA AiHua. Analysis of F-Box Gene Family Based on Salt-Stressed Transcriptome Sequencing in Vicia faba L. [J]. Scientia Agricultura Sinica, 2020, 53(17): 3443-3454.
[15] SHEN JingYuan,TANG MeiLing,YANG QingShan,GAO YaChao,LIU WanHao,CHENG JieShan,ZHANG HongXia,SONG ZhiZhong. Cloning, Expression and Electrophysiological Function Analysis of Potassium Channel Gene VviSKOR in Grape [J]. Scientia Agricultura Sinica, 2020, 53(15): 3158-3168.
Viewed
Full text


Abstract

Cited

  Shared   
  Discussed   
No Suggested Reading articles found!