牦牛Prdm9基因保守区的原核表达及多克隆抗体制备

生物技术通报 ›› 2014, Vol. 0 ›› Issue (3): 155-158.

牦牛Prdm9基因保守区的原核表达及多克隆抗体制备

曾琴, 黄林, 林亚秋, 金素钰, 郑玉才

(西南民族大学生命科学与技术学院，成都 610041)

收稿日期:2013-12-18 出版日期:2014-03-29 发布日期:2014-03-31
作者简介:曾琴，女，硕士，研究方向:动物生化与分子遗传学；E-mail:zengqin886@126.com
基金资助:
国家自然科学基金项目(31240053)，中央高校基本科研业务费资助课题(12NZYTH06)，西南民族大学研究生创新型科研项目(CX2013SZ54)

Prokaryotic Expression and Polyclonal Antibody Preparation of the Conserved Region of Yak Prdm9 Gene

Zeng Qin, Huang Lin, Lin Yaqiu, Jin Suyu, Zheng Yucai

(College of Life Science and Technology，Southwest University For Nationalities，Chengdu 610041)

Received:2013-12-18 Published:2014-03-29 Online:2014-03-31

摘要/Abstract

摘要： 采用逆转录-聚合酶链式反应(RT-PCR)方法，从牦牛睾丸组织获得 Prdm9基因的cDNA的保守区序列，克隆到pMD19-T载体中。经测序确证后，将基因克隆到原核表达载体PET32a(+)，转化 E.coli表达菌BL21(DE3)，以IPTG诱导融合蛋白的高效表达。利用Ni柱亲和层析纯化蛋白，将得到的抗原免疫日本大白兔，获得了1∶4 效价的牦牛PRDM9保守序列的多克隆抗体，可用于后续试验的研究。

关键词: prdm9, 牦牛, 杂交不育, 原核表达, 多克隆抗体

Abstract: The conserved region of Prdm9 cDNA was obtained from yak testis by RT-PCR and cloned into pMD19-T vector．Following sequence confirmation, the cDNA was ligated with pET-32a(+)vector and then transformed into E. coli BL21(DE3)to construct expression strains．The recombinant PRDM9 protein was efficiently expressed in the form of fusion protein through induction with IPTG．After purification by Ni-NTA, the recombinant protein was injected into Japan White rabbits, and polyclonal antibody against the conserved region of PRDM9 of yak was obtained and the antiserum was collected after the titter reach 1∶4, the antibody can be used in the future research．

Key words: Prdm9, Yak, Male infertility hybrid, Prokaryotic expression, Polyclonal antibody

曾琴, 黄林, 林亚秋, 金素钰, 郑玉才. 牦牛Prdm9基因保守区的原核表达及多克隆抗体制备[J]. 生物技术通报, 2014, 0(3): 155-158.

Zeng Qin, Huang Lin, Lin Yaqiu, Jin Suyu, Zheng Yucai. Prokaryotic Expression and Polyclonal Antibody Preparation of the Conserved Region of Yak Prdm9 Gene[J]. Biotechnology Bulletin, 2014, 0(3): 155-158.

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