Identification Of Homologous Recombination Sites During Mice Spermiogenesis

Sperm cells as mammalian sexual reproduction cells,are meiosis product.Recombination of genetic material occurs during meiosis is indispensable for future environmental adaptability and sperm cells genetic materialdiversity.Genetic material phase analysis can distinguish parental origin of alleles,discover meiotic homologous recombination region.Identification of recombination region and its molecu Lar mechanisms research,contribute to further explore some of the causes of genetic diseases,has important reference value for the prevention of genetic disease.This topic contains biological experiments and bioinformatics analysis in two parts.Experimental part include: mouse haploid sperm cells flow sorting;obtain high quality DNA after cleavage separating sperm cells;large fragment sequencing library construction for high-throughput sequencing.Bioinformatics section includes: sequencing data quality control;reference genome sequence comparison;split the data;identification of sequencing fragments' parental origin based on the single nucleotide polymorphisms;enrichment analysis of gene function.Identification the meiosis recombinant chimeric fragments lay the foundation for further study of the relationship between genetic variation and recombination hotspots,and to explore the molecu Lar diversity of the nature of sperm cells.The main findings are as follows:1.Mouse testis cells were stained by Hoechst33342 followed flow sorting successfu Lly isolating haploid sperm cells.The number of sorting reach 106 per 3 male mice;2.Library construction using a long fragment library construction kit to build four libraries are stable.Eventually peak at 900 bp fragment library up and down,the concentration up to 95.2 ng / u L,in line with the quality requirements for library sequencing;3.We get high quality high-throughput sequencing data.High credible single nucleotide mutation sites about 5.17 million,recognizes 299 chimeric fragments;4.Functional enrichment analysis showed that: the presence of the chimeric fragment enriched cellu Lar components include cycle-dependent protein kinase holoenzyme complex,a cell surface molecu Le,transporter Golgi complex and the like;molecu Lar functions have autocrine process,osteoblast differentiation and mesenchymal stem cells canonical Wnt signaling pathway protein receptor,macrophage autophagy down other processes;the biological processes involved in include C-C chemokines and receptor binding activity,and vasoactive intestinal peptide receptor binding PDZ domain activity,glycine aminotransferase activity and the like.These protein or receptor activity affected intracellu Lar material transport,ion channels and various signaling pathways,directly or indirectly involved in cell adhesion,proliferation and differentiation,and tumor cell differentiation and proliferation,and cell-mediated immune processes are also relevant.