秀丽隐杆线虫双突变株系hif-1(ia04);glb-13(tm2825)的制备与鉴定  

Construction and characterization of double mutant strain hif-1(ia04);glb-13(tm2825) in C. elegans

作　　者：石锦平[1] 任长虹[1] 李媛[1] 张成岗[1] 

机构地区：[1]军事医学科学院放射与辐射医学研究所,蛋白质组学国家重点实验室,北京100850

出　　处：《军事医学》2011年第4期253-257,289,共6页Military Medical Sciences

基　　金：国家973计划项目(2006CB504100);国家科技重大专项课题(2009ZX09103-616);国家自然科学基金项目(30771230;30600365);中国博士后科学基金特别资助基金(200801495)

摘　　要：目的以秀丽隐杆线虫为模式生物研究珠蛋白(globin)13(glb-13)的生理作用,以回交2次的glb-13(tm2825)突变株系和hif-1(ia04)构建线虫双突变株系hif-1(ia04);glb-13(tm2825)。方法 glb-13(tm2825)与野生型N2株系交配获得glb-13(tm2825)的雄虫,再与N2的雌雄同体线虫进行交配,回交2次后,通过单只线虫基因组PCR法鉴定出交配后的纯合体。使用hif-1(ia04)和野生型线虫N2进行交配得到hif-1(ia04)的雄虫,然后再与glb-13(tm2825)雌雄同体线虫交配,通过单只线虫基因组PCR法鉴定出交配后的纯合体。结果获得hif-1(ia04);glb-13(tm2825)双突变株系。结论通过线虫交配回交可以使突变株系线虫保持稳定性状,而突变株系之间的交配可以制备出双突变株系,为深入开展glb-13的功能研究奠定了物质基础。Objective To investigate the physiological function of the glb-13 gene,to backcross glb-13（tm2825） and to construct double mutant strain hif-1（ia04）;glb-13（tm2825） in Caenorhabditis elegans.Methods The glb-13（tm2825） mutant male strain was obtained by mating the glb-13（tm2825） mutant hermaphrodite with the male N2 strain,and then with the N2 hermaphrodite.It was then backcrossed twice,and the homozygote was identified by single C.elegans genome PCR.The male hif-1（ia04） mutant strain,which was obtained by mating the hif-1（ia04） mutant hermaphrodite with wild-type N2 strain,was further mated with the glb-13（tm2825） mutant hermaphrodite.The homozygote was also identified by single C.elegans genome PCR.Results The backcrossed mutant strain glb-13（tm2825） and the homozygotes hif-1（ia04）;glb-13（tm2825） double mutant were obtained.Conclusion Backcross can maintain the mutant′s genetic stability while mutant mating can construct a double mutant.This finding will facilitate further functional studies of the glb-13 gene.

关 键 词：秀丽隐杆线虫 交配 回交 突变 珠蛋白 两性畸形 活性氧类 低氧诱导因子1 

分 类 号：R383.1[医药卫生—医学寄生虫学]