The Hu-IFN-αgene,which was transducted into downstream promoter of β-actin gene of common carp（Cyprinus carpio）,was recombined by DNA recombination technology.These recombined genes were injected into 1-2 cell fer...The Hu-IFN-αgene,which was transducted into downstream promoter of β-actin gene of common carp（Cyprinus carpio）,was recombined by DNA recombination technology.These recombined genes were injected into 1-2 cell fertilized eggs of grass carp（Ctenophatyngodon idellus） by micro-injection technology,we gained transgenetic fish by molecular detection methods.In order to analyse the genetic expression of tran-Hu-IFN-α gene gynogensis F1,which male individualization were gained by raising methyl-testosterone,molecular genetic marker technology was used.In our research,30 random primers were picked out from 48 and were used into RAPD-PCR,the result indicated that 1 169 clear,steady and repeated DNA finger printing bands were achieved.On the basis of gentic distance matrix among tran-Hu-IFN-α gene gynogenesis F1 group,the genetic relationship of gynogenesis F1 were analysed by UPGMA,the results showed the genetic patterns are close between the 3# male gynogenesis F1 and the the 23# female of gynogenesis F1,5# and 27#,2# and 28#,2# and 30#.The data indicated that these group could be served as parent of tran-Hu-IFN-α grass carp（Ctenophatyngodon idellus） pure line.展开更多
文摘目的为HLA和CD36复合抗体所致血小板输注无效(PTR)患者寻求相合或相容的供者血小板输注。方法采用ELISA方法检测PTR患者的血小板抗体及HLA⁃Ⅰ类抗体特异性;运用MATCH IT!和HLA Matchmaker软件,分析患者HLA⁃Ⅰ类抗体特异性及相应抗原决定簇(epitopes);采用HLA⁃SSO分型技术,获得供、患者的HLA基因型;根据HLA⁃I类抗原交叉反应组(CREG)或HLA表位配型(Eplet)策略,寻找与PTR患者相合或相容的供者血小板;通过血小板抗原单克隆抗体特异性免疫固定检测技术(MAIPA)和血小板免疫荧光流式检测(PIFT)鉴定匹配程度;最后,通过血小板计数纠正增加指数(CCI)评估血小板输注效果。结果2名PTR患者体内检测到针对HLA⁃Ⅰ类和CD36的复合抗体,且CD36流式表型为Ⅰ型缺失;抗体特异性检测结果显示,患者1和患者2的血清中存在高频的HLA⁃Ⅰ类抗体,PRA分别为56%(54/96)和53%(51/96);运用HLA的CREG和Eplet配型策略,在CD36缺失供者库分别筛选到与患者1匹配等级C的供者1名,与患者2匹配等级D的供者1名,且选择的供者均规避患者HLA⁃Ⅰ类抗体表位所针对的抗原;MAIPA和PIFT结果亦证实患者与供者血小板无免疫反应,且患者2输注相容血小板24 h CCI>4.5。结论针对HLA和CD36复合抗体所致PTR患者,可联合运用血清学交叉配型、HLA⁃CREG及Eplet配型策略,选择CD36缺失,规避HLA⁃Ⅰ类抗体对应抗原且HLA表位匹配的供者血小板。
文摘The Hu-IFN-αgene,which was transducted into downstream promoter of β-actin gene of common carp（Cyprinus carpio）,was recombined by DNA recombination technology.These recombined genes were injected into 1-2 cell fertilized eggs of grass carp（Ctenophatyngodon idellus） by micro-injection technology,we gained transgenetic fish by molecular detection methods.In order to analyse the genetic expression of tran-Hu-IFN-α gene gynogensis F1,which male individualization were gained by raising methyl-testosterone,molecular genetic marker technology was used.In our research,30 random primers were picked out from 48 and were used into RAPD-PCR,the result indicated that 1 169 clear,steady and repeated DNA finger printing bands were achieved.On the basis of gentic distance matrix among tran-Hu-IFN-α gene gynogenesis F1 group,the genetic relationship of gynogenesis F1 were analysed by UPGMA,the results showed the genetic patterns are close between the 3# male gynogenesis F1 and the the 23# female of gynogenesis F1,5# and 27#,2# and 28#,2# and 30#.The data indicated that these group could be served as parent of tran-Hu-IFN-α grass carp（Ctenophatyngodon idellus） pure line.
