Objective To try making huZP3a^22-176 and huZP3b^177-348 polypeptides （representing an intact huZP^322-348 protein without its N-terminal signal peptide and C-terminal transmembrane domain ） express in E. coli at a ...Objective To try making huZP3a^22-176 and huZP3b^177-348 polypeptides （representing an intact huZP^322-348 protein without its N-terminal signal peptide and C-terminal transmembrane domain ） express in E. coli at a higher level Methods The cDNAs encoding huZP3a and huZP3b were obtained with PCR method. The pBV221 plasmid was used to construct thermo-inducible recombinant expression vector. Purification of two target expression products employed an improved method of preparative gel polyacrylamide gel electrophoresis. Results Two polypeptides of recombinant huZP3a （rhuZP3a） and recombinant huZP3b （rhuZP3b） were all expressed respectively in an E. coli BL21（DE3）pLysS strain at a higher level, which were recognized by two specific polyclonal antisera in Western blotting test which recognize a linear B cell epitope present in rhuZP3a or rhuZP3b respectively. Using the shake-flask method, approximately 5 mg of rhuZP3a and rhuZP3b with more than 95% relative homogeneity were harvested from 1 L culture respectively. Conclusion The availability of two rhuZP3 polypeptides will help in detecting the immunogenicities of rhuZP3a and rhuZP3b through animal experiments and confirming the function domain of non-glycosylated huZP3 to induce acrosome reaction in vitro.展开更多
Job recruitment livestreams are boosting employment by instantly connecting companies with the nationwide talent pool.But as fraudsters lure unsuspecting blue-collar job seekers with false listings,experts are calling...Job recruitment livestreams are boosting employment by instantly connecting companies with the nationwide talent pool.But as fraudsters lure unsuspecting blue-collar job seekers with false listings,experts are calling for greater regulation.展开更多
Once the sperm enters into the female reproductive tract,they undergo an important process called capacitation,which involves a series of physiological and biochemical changes that ensure fertilization.During capacita...Once the sperm enters into the female reproductive tract,they undergo an important process called capacitation,which involves a series of physiological and biochemical changes that ensure fertilization.During capacitation,calcium ions(Ca^(2+))increase the bending amplitude of a sperm’s flagellum.This causes hyperactivation,which is manifested as the asymmetrical beating of the flagellum.Tyrosine phosphorylation is also a cause for human sperm hyperactivation;asthenospermia—that is,reduced sperm motility—occurs when the sperm are incapable of hyperactivation owing to impaired tyrosine phosphorylation.[1]Hyperactivated motility enables the sperm to migrate and penetrate the cumulus cells and zona pellucida surrounding the oocyte,leading to fertilization.Therefore,the hyperactivated motility of sperm is critical for male reproductive ability,and the molecular mechanism underlying human sperm hyperactivation requires discussion.展开更多
基金This work was supported by grant (No. 03JG05014) from the Population Family Planning Commission ofShanghai. China and the Medical and Health Science Research Foundation of Zhejiang Province(No. 2004A002)
文摘Objective To try making huZP3a^22-176 and huZP3b^177-348 polypeptides （representing an intact huZP^322-348 protein without its N-terminal signal peptide and C-terminal transmembrane domain ） express in E. coli at a higher level Methods The cDNAs encoding huZP3a and huZP3b were obtained with PCR method. The pBV221 plasmid was used to construct thermo-inducible recombinant expression vector. Purification of two target expression products employed an improved method of preparative gel polyacrylamide gel electrophoresis. Results Two polypeptides of recombinant huZP3a （rhuZP3a） and recombinant huZP3b （rhuZP3b） were all expressed respectively in an E. coli BL21（DE3）pLysS strain at a higher level, which were recognized by two specific polyclonal antisera in Western blotting test which recognize a linear B cell epitope present in rhuZP3a or rhuZP3b respectively. Using the shake-flask method, approximately 5 mg of rhuZP3a and rhuZP3b with more than 95% relative homogeneity were harvested from 1 L culture respectively. Conclusion The availability of two rhuZP3 polypeptides will help in detecting the immunogenicities of rhuZP3a and rhuZP3b through animal experiments and confirming the function domain of non-glycosylated huZP3 to induce acrosome reaction in vitro.
文摘Job recruitment livestreams are boosting employment by instantly connecting companies with the nationwide talent pool.But as fraudsters lure unsuspecting blue-collar job seekers with false listings,experts are calling for greater regulation.
基金supported by the Natural Science Foundation of China(Nos.81771647,81801525,81571426,and 81170554)Natural Science Foundation of Zhejiang Province(No.LQ17H040004)+2 种基金Health Sci&Tech Plan Project of Zhejiang Province(Nos.2018KY039 and 2019KY363)Special Project for the Research Institutions of Zhejiang Province(Nos.YS2021014 and C11920D-04)Zhejiang Provincial Program for the Cultivation of High-level Innovative Health Talents,and the Start-up Fund of Hangzhou Medical College(No.C11904Q-04)。
文摘Once the sperm enters into the female reproductive tract,they undergo an important process called capacitation,which involves a series of physiological and biochemical changes that ensure fertilization.During capacitation,calcium ions(Ca^(2+))increase the bending amplitude of a sperm’s flagellum.This causes hyperactivation,which is manifested as the asymmetrical beating of the flagellum.Tyrosine phosphorylation is also a cause for human sperm hyperactivation;asthenospermia—that is,reduced sperm motility—occurs when the sperm are incapable of hyperactivation owing to impaired tyrosine phosphorylation.[1]Hyperactivated motility enables the sperm to migrate and penetrate the cumulus cells and zona pellucida surrounding the oocyte,leading to fertilization.Therefore,the hyperactivated motility of sperm is critical for male reproductive ability,and the molecular mechanism underlying human sperm hyperactivation requires discussion.
