也木勒羊抑制素α亚基基因克隆及其真核表达

doi:10.16431/j.cnki.1671-7236.2015.06.003

Abstract

Abstract: To clone the Yemule aries inhibin α (INHα) subunit gene and express its recombinant protein,total RNA was extracted from Yemule aries ovary tissue by Trizol,and a pair of primer was designed according to the published aries INHα subunit gene sequence (GenBank No.XM_004004955.1). INHα subunit gene coding sequence was amplified by RT-PCR.The mature peptide cDNA was further amplified and cloned into the Scal Ⅰ and EcoR Ⅰ sites of the pEGFP-N1 expressing vector to generate the recombinant plasmid pEGFP-INHα.The plasmid was transformed into DH5α,after appraisal,sequencing,proves that successful build INHα subunit gene eukaryotic expression vector.The sequence result was compared with homologous sequences from animals including human,gorilla,cow and other animals alpha subunit of INHα subunit gene sequence homology comparison analysis.Sequence alignment showed that ranks of their similarities were above 50%,proved that INHα subunit gene was highly conservative.By RT-PCR and Western blotting method verified the eukaryotic expression vector of statin INHα subunit gene expression and protein expression of success in BHK cells.

Key words: Yemule aries; folliculostatin; eukaryotic expression; homology analysis

CLC Number:

HUANG Zeng-wen, Muerzhati, WureliHazi·Hazihan. Cloning and Eukaryotic Expression of Inhibin alpha Gene in Yemule Aries[J]. , 2015, 42(6): 1340-1347.

References

[1] Medan M S,Arai K Y,Watanabe G,et al.Inhibin:Regulation of reproductive function and practical use in females[J].Animal Science Journal,2007,78(1):16-27.

[2] Meldi K M,Gaconnet G A,Mayo K E.DNA methylation and histone modifications are associated with repression of the inhibin a promoterin the rat corpus luteum[J].Endocrinology,2013,153(10):4905.

[3] Knight P G,Glister C.Potential local regulatory functions of inhibins,activins and follistatin in the ovary[J].Reproduction,2001,121:503-512.

[4] Li C,Zhu Y L,Xue J H,et al.Immunization against inhibin enhances both embryo quantity and quality in Holstein heifers after superovulation and insemination with sex-sorted semen[J].Theriogenology,2009,71(6):1011-1017.

[5] Vale W,Rivier C,Hsueh A,et al.Chemical and biological characterization of the inhibin family of protein hormones[J].Recent Prog Horm Res,1988,44:1-34.

[6] Medan M S,Akagi S,Kaneko H,et al.Effects of reimmunization of heifers against inhibin on hormonal profiles and ovulation rate [J].Reproduction,2004,128:475-482.

[7] Ishigame H,Medan M S,Watanabe G,et al.A new alternative method for superovulation using passive immunization against inhibin in adult rats[J].Biol Reprod,2004,71:236-243.

[8] Kusinan T,Meyer R L,Carlson K M,et al.Passive immunization of ewes against an inhibin like peptide increases follicle stimulating hormone concentrations,ovulation rate,and proliflcacy in spring-mated ewes[J].J Anim Sci,1995,73(5):1433-1439.

[9] Fray M D,Wrathall J H,Knight P G.Active immunization against inhibin promotes a recurrent increase in litter size in sheep[J].Vet Rec,1994,134(1):19-20.

[10] Henderson K M,Franchimont P,Lecomte-Yerna M J,et al.Increase in ovulation rate afer active immunization of sheep with inhibin partially purified from bovine follicular fluid[J].J Endocrinol,1984,102(3):305-309.

[11] Wheaton J E,Meyer R L,Jones R H,et al.Effects of passive immunization using antibody against an alpha-inhibin peptide on follicle-stimulating hormone concentrations and litter size in sows [J].Theriogenology,1998,49(4):813-822.

[12] Morris D G,Mcdermott M G,Diskln M G,et al.Effect of immunization against synthetic peptide sequences of bovine inhibin alpha-subunit on ovulation rate and twin-calving rate in heifers[J].J Reprod Fertil,1993,97(1):255-261.

[13] Al-Obaidi S A,Bindon B M,Hillard M A,et al.Reproductive characteristics of lambs actively immunized early in life with inhibin-enriched preparations from follicular fluid of cows[J].J Reprod Fertil,1987,81(2):403-414.

[14] Bame J H,Dalton J C,Degelos S D,et al.Effect of long-term immunization against inhibin on sperm output in bulls[J].Biol Reprod,1999,60(6):1360-1366.

[15] 决肯·阿尼瓦什,库木尼斯汗·加汗,依明·苏来曼,等.新疆额敏县也木勒白羊(暂名)羔羊的屠宰测定[J].新疆农业科学,2006,43(5):443-445.

[16] Ying S Y.Inhibins,activins,and follistatins:Gonadal proteins modulating the secretion of FSH[J].Endocrine Reviews,1988,9:267-293.

[17] Vale W,Rivier J.Purification and characterization of a FSH releasing protein from porcine ovarian follicular fluid[J].Nature,1986,321:776-779.

[18] Findlay J K.An update on the roles of inhibin,activin,and follistatin as local regularors of folliculogenesis[J].Biol Reprod,1993,48(1):15-23.

[19] Ling N,Ying S Y,Ueno N.et al.Isolation and partial characterization of a Mr 32,000 protein with inhibin activity from porcine follicular fluid[J].Proc Nat Acad Sci USA,1985,82(21):7217-7221.

[20] 田锦,李志民,傅衍,等.抑制素、活化素和卵泡素研究进展[J].动物医学进展,2008,29(12):77-81.

[21] Chen Y,MuLler J D,Ruan Q,et al.Molecular brightness charac-terization of EGFP in vivo by fluorescence fluctuation spectroscopy[J].Biophys,2002,82(1):133-134.

[22] 邓继贤,杨秀荣,韦英明,等.沼泽型水牛卵泡抑素基因的真核表达研究[J].中国畜牧兽医,2011,38(9):57-60.

Cloning and Eukaryotic Expression of Inhibin alpha Gene in Yemule Aries

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	PAN Pengcheng, LEI Zongquan, LI Xian, HU Xiangyun, QIN Qiantao, QIN Zhaoxian, GUAN Zhihui, CHEN Baojian, XIE Bingkun. Bioinformatics Analysis,Eukaryotic Expression Vector Construction and Tissue Expression of MYL2 Gene in Luchuan Pigs [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(5): 1796-1806.
[2]	BAI Shaocheng, ZHOU Juan, JIN Rongshuai, WANG Fan, LU Tingting, TANG Xianwei, ZHAO Bohao, WU Xinsheng, CHEN Yang. Cloning and Bioinformatics Analysis of CYP11A1 Gene in New Zealand White Rabbits and Its Effects on Reproduction-related Genes [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(7): 2484-2496.
[3]	ZOU Jianwei, ZOU Juhong, SHEN Yujian, WEI Yi, ZHANG Sanbao, LIAN Zitong, XU Jianjian, SONG Ying, HUANG Yanna, WEI Yingming, JIANG Qinyang, ZHENG Zihua. Bioinformatics Analysis, Construction of Eukaryotic Expression Vector and Expression of LMCD1 Gene in Nubian Goats [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(6): 2033-2042.
[4]	ZHANG Xinjie, XUE Shaohua, LYU Zixin, HUANG Xirong, CHEN Yuxuan, FAN Kewei, YANG Xiaoyan, DAI Ailing. Investigation of Co-infection of PPV7 and PCV2 and Genetic Variation Analysis for PPV7 Cap Gene in Fujian and Guangdong [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(6): 2291-2297.
[5]	DONG Yuan, YU Hongxu, CHEN Guoliang, BAO Dongbo, CUI Jiarui, LIU Lei, WANG Huiyan. Construction of Eukaryotic Expression Vector of Bovine Pregnancy-associated Glycoprotein 19 and Its Bioinformatics Analysis [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(4): 1233-1243.
[6]	PANG Feng, LONG Qinqin, LIANG Shaobo, CHENG Hongsong, CHENG Zhentao. Bioinformatics Analysis,Eukaryotic Expression and Subcellular Localization of Orf Virus ORFV114 Protein [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(11): 4150-4158.
[7]	CAO Yingying, LI Huijun, LI Baoying, LIANG Liang, LENG Jing, TANG Haibo. Expression of ISG15 Protein of Tupaia belangeri yaoshanensis and Preparation of Its Polyclonal Antibody [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(1): 273-282.
[8]	HU Huanyi, WANG Jianzhong, LIU Changjin, LIN Min, LIU Xiaolan, WEI Huangsiwu, DENG Shunzhou. Eukaryotic Expression of Green Fluorescent Protein and Preparation of Its Monoclonal Antibody [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(1): 328-337.
[9]	QIAO Lianjiang, CHEN Chen, ZHANG Ru, CUI Liheng, XU Li, YANG Yanling. Construction of Eukaryotic Expression Vectors and Bioinformatics Analysis of Rhoa and Ptgs2 Genes in RAW264.7 Cells [J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(7): 2313-2323.
[10]	ZOU Juhong, SHEN Yujian, GAO Xiaotong, HUANG Yanna, JIANG Qinyang. Cloning and Analysis of FTO Gene of Nubian Goat and Construction of Eukaryotic Expression Vector [J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(7): 2342-2348.
[11]	JIAO Wenqiang, LIU Yunchao, XING Guangxu, CHEN Pengju, XU Yindi, WANG Zhifang, ZHANG Qingxian, XU Feng. Bioinformatics Analysis and Eukaryotic Expression of Chlorocebus sabaeus Glucose Regulatory Protein 78 [J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(4): 1170-1178.
[12]	ZHAO Hongcong, GAO Dengke, JIANG Haizhen, LI Yating, LI Dan, MA Bairong, JIN Yaping, CHEN Huatao. Construction of Eukaryotic Expression Vector and Bioinformatics Analysis of Circadian CLOCK Gene in Goats [J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(12): 4327-4338.
[13]	PAN Pengcheng, WEN Binhua, XIE Wan, JIANG Changjin, JIAO Di, CHEN Baojian, GUAN Zhihui, XIE Bingkun. Sequence Analysis,Eukaryotic Expression Vector Construction and Tissue Expression Analysis of PDK4 Gene in Luchuan Pigs [J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(8): 2337-2347.
[14]	WANG Zhaoyang, JIANG Yajun, LIU Xueting, LIN Weidong, XIN Ting, HOU Shaohua, GUO Xiaoyu, ZHU Hongfei, JIA Hong. Construction and Screening of Phage Library Against VHH Antibody of Canine Distemper Virus [J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(6): 1685-1693.
[15]	YAN Xiukui, HAN Zhixiao, DU Qian, WANG Wei, ZHU Yuanzhi, XIN Jialiang, YI Chizhe, BI Jingshan, SUN Wenchao, ZHENG Min. Detection and Sequence Analysis of Canine Circovirus in Chongqing [J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(10): 3069-3077.