权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Introduction and expression of foreign gene in fish

外源基因在鱼类中的导入及表达

基本信息

批准号：
01480080
负责人：
AOKI Takashi
金额：
$ 3.97万
依托单位：
Faculty of Agriculture, Miyazaki University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (B)
财政年份：
1989
资助国家：
日本
起止时间：
1989 至 1990
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-01480080/
关键词：
fish globin gene transgenic fish expression of foreign gene in fish promoter region chloramphenicol acetyltransferase gene 5'-flanking and 5'-untranslated regions fish cell line transfection 魚類のグロビン遺伝子トランスジェニックフィッシュ魚類における外来遺伝子の発現プ

项目摘要

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.

克隆了黄尾和鲤鱼类α珠蛋白基因。在鱼类珠蛋白基因的5'侧翼区发现了-100区、CCAAT盒和TATA盒的启动子元件。帽位点位于黄尾鱼类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。将氯霉素乙酰转移酶（CAT）基因与黄尾鱼的5 '侧翼区和5'非翻译区连接到载体质粒中。将重组质粒导入三种鱼类组织培养系中。在转染了CAT基因5 ′侧翼区和5 ′非翻译区重组质粒的鱼组织培养细胞系中检测到了明显的CAT活性。将鲤鱼α-珠蛋白基因（CalphaG）显微注射到受精后的虹鳟鱼卵细胞质中，用谷胱甘肽（GSH）处理受精后的1细胞期虹鳟鱼卵细胞质中，观察到5 '侧翼区和5'非翻译区具有启动子活性。约67%的卵孵化并正常生长。从130日龄鱼和10 ~ 12月龄鱼中检测到CalphaG基因。整合和非整合的细胞在宿主鱼中镶嵌共存。将CalphaG基因导入虹鳟鱼（Oncorhynchus mykiss）后，可将其遗传给后代。