Introduction and expression of foreign gene in fish

克隆了黄尾鱼和鲤鱼的类α珠蛋白基因。-100区、CCAAT盒和TATA盒的启动子元件位于鱼类珠蛋白基因的5‘侧翼区。该帽区位于黄尾鱼类α珠蛋白基因ATG起始密码子上游46个碱基对，鲤鱼类α珠蛋白基因ATG序列上游48个核苷酸。将黄尾病毒的5‘端和5’端非翻译区与载体载体中的氯霉素乙酰转移酶(CAT)基因连接。将重组载体导入三种鱼类组织培养系中。在5‘-侧翼和5’-非翻译区-CAT重组质粒转染的鱼类组织培养细胞系中检测到显著的CAT活性。这一结果表明，鱼类珠蛋白基因的5‘-侧翼区和5’-非翻译区具有启动子活性，在鱼类组织培养细胞的基因表达调控中起重要作用。约67%的显微注射卵孵化出来并正常生长。从130日龄鱼和10~12月龄鱼中检测到CalphaG基因。在寄主鱼类体内，CalphaG整合细胞和非整合细胞嵌合共存。将CalphaG基因导入虹鳟鱼(Oncorhynchus Mykises)，可以遗传给后代。

The alpha-like globin genes from yellowtail (Seriola quinqueradiata) and carp (Cyprinus carpio) were cloned. The promoter elements of the -100 region, CCAAT box, and TATA box were found in the 5' -flanking region of fish globin genes. The cap site is located 46 base pairs upstream from ATG initiationcodon of the alpha-like globin gene of yellowtail and is 48 nucleotides upstream from the ATG sequences of the alpha-like globin gene of carp. The 5'-flanking and 5'-untranslated regions of yellowtail were ligated with chloramphenicol acetyltransferase (CAT) gene in vector plasmid. The recombinant plasmids were introduced into three kinds of fish tissue culture lines. The significantly CAT activity was detected in the fish tissue culture cell lines transfected with the 5'-flanking and 5'-untranslated regions-CAT recombinant plasmids. This result suggests the 5'-flanking region and 5'-untranslated regions from fish globin gene possess the promoter activity and play an important role in regulation of gene expression in fish tissue culture cells.Carp alpha-globin gene (CalphaG) was microinjected into the cytoplasm of fertilized rainbow trout eggs at 1 cell stage treated with glutathione. About 67% of microinjected eggs hatched out and grew normally. CalphaG gene was detected from 130-day-old fish and 10 to 12-month-old fish. CalphaG-integrated and non-integrated cells coexisted mosaically in host fish. The CalphaG gene introduced into rainbow trout (Oncorhynchus mykiss) could be transmitted to their offspring.