首页|Prokaryotic Expression, Ascitic Polyclonal Antibody Preparation and Identification of Cashmere Goat Izumo1
Prokaryotic Expression, Ascitic Polyclonal Antibody Preparation and Identification of Cashmere Goat Izumo1
扫码查看
点击上方二维码区域，可以放大扫码查看
原文链接
国家科技期刊平台
NETL
NSTL
万方数据
Izumol is a novel member of the immunoglobulin superfamily locating on sperm, and is indispensable for sperm-egg fusion. According to its immunoglobulin-like domain in the extracellular region, Izumol was fractionated into 6 fragments (F0-F5) which were ligated with pGEX-4Tl to construct the prokaryotic expression vectors pGEX-Fn. The recombinant plasmids were transformed into Escherichia coli BL21 (DE3) and the GST-Fn fusion proteins were expressed successfully by induction with IPTG. GST-FO, a recombinant fusion protein of GST with the full length of extracellular region of mature cashmere goat Izumol, was purified by polyacrylamide gel slicing method and was used as an antigen to immunize the Kunming mouse to generate anti-GST-Izumo1 ascetic polyclonal antibody with intraperitoneal injection of SI80 cells. Subsequently, the anti-GST-Izumol polyclonal antibody was purified with miscellaneous antigen by glutaraldehyde cross-linking method. Western blotting analysis showed that the purified ascetic polyclonal antibody had high affinity to all 6 GST-Izumo1 fragment fusion proteins. Immunohistochemical analysis with this antibody displayed that the cashmere goat Izumol proteins were at the equatorial segment of sperm head surface. These results indicate that this polyclonal antibody has high specificity and lays the foundations for further study on the expression pattern of Izumol in cashmere goat testis and binding abilities of each extra-membrane fragment of Izumol to the egg surface.
LIU Zhi-da、XING Wan-jin、WANG Lian-qing、L(U) Li-xia
展开 >
Key Laboratory of Mammalian Reproduction Biology and Technology, Ministry of Education/School of Life Sciences, Inner Mongolia University, Hohhot 010021, P.R.China
Natural Science Foundation of Inner Mongolia Autonomous Region, ChinaNational Training Foundation for Talents of Basic Sciences, China
国家科技期刊平台
NSTL
万方数据
