新霉素抗性基因在家蚕中的插入和表达  

Transformation of Neomycin Resistance Gene ( neo R) into Silkworm ( Bombyx mori.L. )

作　　者：陈秀[1,2] 赵昀[1,2] 张峰[1,2] 彭卫平 冯晓黎 黄君霆[1,2] 陆长德[1,2] 

机构地区：[1]中国农业科学院蚕业研究所 [2]中国科学院上海生物化学研究所

出　　处：《生物化学与生物物理学报》1999年第1期90-92,共3页

摘　　要：构建含新霉素抗性基因（ｎｅｏｍｙｃｉｎｒｅｓｉｓｔａｎｃｅｇｅｎｅ，ｎｅｏＲ）的重组质粒ｐＦＮ，经ＨｉｎｄＩＩ酶切后，用基因枪将ＤＮＡ片段导入家蚕早期受精卵中（Ｇ０代）。孵化的Ｇ１、Ｇ２代蚁蚕均经含新霉素的人工饲料添食２４ｈ后，筛选出新霉素抗性的个体（能正常生长发育的）改为桑叶饲养。于Ｇ２代的５龄第二天从后部丝腺抽提总ＤＮＡ，再以ｎｅｏＲ的ｃＤＮＡ为探针进行Ｓｏｕｔｈｅｒｎ杂交检测。结果表明ｎｅｏＲ基因已转入家蚕ＤＮＡ中，获得了含ｎｅｏＲ的转基因蚕。Neomycin resistance gene ( neo R) flanked by 5′ and 3′ regions of fibroin H chain gene of silkworm ( Bombyx mori.L. ) was transferred into eggs of silkworm by gene gun in the early period of fertilization. The larvae were fed with an artificial diet containing neomycin in early 24 hours post transfettion, and some of them survived. The neo R encoding sequence in G 2 generation derived from the survivals was detected by Southern blotting. The results indicated that neo R could be used as a selective marker for studies on transgenic silkworm.

关 键 词：家蚕 筛选标记 neo^R 转基因蚕 

分 类 号：S882.2[农业科学—特种经济动物饲养]