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| Expression of Ovine Myostatin Gene and Construction and Identification of Nanobody Library Against Recombinant MSTN |
| LI You-jian1, ZHANG guo-qi2, Gou ji-xing2, CHEN xin-kai1, DOU Xiao-xia1, CHEN Chuang-fu2, SHENG Jin-liang1 |
1. College of Animal Science and Technology;
2. Key Laboratory of Xinjiang Endemic and Ethnic Disease, Shihezi University, Shihezi 832003, China |
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Abstract The ovine myostatin gene was subcloned into the pET-32a(+),and transformed into E.coli BL21.The MSTN recombinant fusion protein was purified and the healthy Bactrian camel was selected for immunization with the MSTN protein.Total RNA was extracted from peripheral lymphocytes for the amplification of VHH fragments by RT-PCR.Primers designed according to the single domain gene sequences of the antibodies and the variable camel IgG2, IgG3 genes were amplified by Nested PCR.PCR products were then purified and inserted into phage mid vector pCANTAB5 E. The VHH antibody gene library was obtained by electroporating recombinant pCANTAB5E VHH vectors into E. coli TG 1 cells.The first set of antibody gene library was composed with 9.5×105 individual colones after cloning VHH genes and the library we possessed high diversity and good capacity,which provides a platform for panning VHH camilid antibody aimed at ovine MSTN.
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Received: 14 July 2014
Published: 25 September 2014
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