Cloning and expression of mouse sperm protein Sp17 and its immunological effects
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Cloning and expression of mouse sperm protein Sp17 and its immunological effects
Journal of Army Medical UniversityVol. 30, Issue 24, Pages: 2281-2284(2008)
作者机构:
1. 第三军医大学:西南医院妇产科,基础医学部全军免疫学研究所
2. 解放军总医院第二临床部妇产科
作者简介:
基金信息:
DOI:
CLC:R341;R392.2;R321.1
Published:2008,
Published Online:30 December 2008,
稿件说明:
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ZHANG QIAO-YU, LIANG ZHI-QING, LI YU-YAN, et al. Cloning and expression of mouse sperm protein Sp17 and its immunological effects. [J]. Journal of army medical university, 2008, 30(24): 2281-2284.
DOI:
ZHANG QIAO-YU, LIANG ZHI-QING, LI YU-YAN, et al. Cloning and expression of mouse sperm protein Sp17 and its immunological effects. [J]. Journal of army medical university, 2008, 30(24): 2281-2284.DOI:
Cloning and expression of mouse sperm protein Sp17 and its immunological effects
Objective To clone mouse sperm Sp17 gene and fulfill its expression in
E. coli
BL21(DE3). Methods The coding sequence of mouse protein Sp17 was amplified from mouse testis RNA through reverse transcription-polymerase chain reaction (RT-PCR)
and subsequently cloned into pET-28a (+). Recombinant Sp17 was then expressed in
E. coli
BL21 (DE3) with induction of IPTG and analyzed by SDS-PAGE and Western blotting. The mice were immunized intranasally with the recombinant proteins and the titers of specific antibodies IgG in serum were dete
cted. Results Sequencing and restriction digestion of the recombinant plasmid demonstrated that the coding sequence of mouse protein Sp17 was successfully cloned. SDS-PAGE and Western blot analysis showed that a recombinant protein with molecular weight of 24 000
which is corresponding to the theoretical molecular weight of Sp17 protein
existed in the extract of
E. coli
BL21 (DE3) cells. After immunization
Sp17-specific antibody IgG response was present in the mice. Conclusion The coding sequence of mouse Sp17 was cloned into pET-28a (+) plasmid vector
and high level of recombinant Sp17 protein expressed in
E. coli
BL21 (DE3) was obtained. The recombinant Sp17 protein can induce Sp17-specific antibody IgG response.