男性不育患者精液来源的金黄色葡萄球菌精子制动相关基因研究

A gene related to inhibition of sperm mobility in Staphylococcus aureus from the sperm of infertile men

目的:分析分离自男性不育症患者精液的精子制动阳性金黄色葡萄球菌的全基因组信息,发掘可能编码导致精子制动的蛋白和基因。方法:收集分离男性不育症患者精液中的金黄色葡萄球菌(共22株),并根据其对精子的制动能力分为精子制动阳性组与制动阴性组,随后选出各组最具有代表性的菌株MJ015(制动阳性株)与MJ163(制动阴性株),分别提取其DNA进行全基因组测序,对获得的测序结果进行全基因组序列拼接并提交NCBI进行注释,MJ015的NCBI登录号为CP038183,MJ163的NCBI登录号为CP038229,同时使用BRIG软件进行BLAST比对并制作圈图,使用Artemis软件套装对MJ015与MJ163进行详细比对以寻找目标基因。结果:MJ015的染色体全基因组序列总长为2784836 bp,含有2个质粒,MJ163的染色体全基因组序列总长为2746673 bp,包括1个质粒,MJ015和MJ163的GC含量分别为32.13%、32.08%,MJ015和MJ163分别注释有2921个和2844个基因,且经过比对发现MJ015与MJ163的全基因组序列有约130 kb的序列未能比对上,在这些差异序列中所包含的SAK基因,其表达产物经实验证实为这两株菌表达的差异蛋白。结论:经序列分析与实验验证,金黄色葡萄球菌MJ015基因组上的SAK基因可能是使精子制动的关键基因,其表达的产物葡激酶对精子制动的能力强弱的影响有待进一步研究证实。

Objective:To analyze the whole genome sequences of Staphylococcus aureus strains from the sperm of infertile males and identify the gene which may induce the inhibition of sperm motility(ISM).Methods:Twenty-two Staphylococcus aureus strains were isolated from the sperm of infertile males in the First Hospital Affiliated to Wenzhou Medical University and,according to the ability of ISM,divided into an ISM and a non-ISM group.Two strains most representative of the biological function of each group were selected,namely MJ015 from the ISM and MJ163 from the non-ISM group,and DNA extracted from them for whole genome sequencing.The data obtained were subjected to whole-genome sequence assembly and submitted to NCBI for annotation,with the accession number of CP038183 for MJ015 and CP038229 for MJ163.The whole genome sequences of MJ015 and MJ163 were compared in full detail using BRIG and Artemis software suite to identify the target gene.Results:The whole genome sequence of MJ015 was 2784836 bp in length,containing 2 plasmids,and that of MJ163 was 2746673,containing 1 plasmid,each with a 32.13%,32.08%content of guanine-cytosine(GC),and annotated with 2921 and 2844 genes respectively.Comparison between the whole genome sequences of MJ015 and MJ163 revealed an almost 130 kb gap,in which a gene named sak was found to express a potential serum inhibition factor,whose transcription product was proved to be a differentially expressed protein in the two strains.Conclusion:The gene sak in MJ015 may play a key role in the inhibition of sperm mobility,but the inhibition intensity of its transcription product staphylococcus kinase has to be further studied.