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pRSV-虹鳟鱼生长激素cDNA在鲤(鲤属,林奈)中的基因转移、表达及遗传

Gene transfer, expression and inheritance of pRSV-rainbow trout-GH cDNA in the common carp, Cyprinus carpio (Linnaeus).

作者信息

Zhang P J, Hayat M, Joyce C, Gonzalez-Villaseñor L I, Lin C M, Dunham R A, Chen T T, Powers D A

机构信息

Hopkins Marine Station, Stanford University, Pacific Grove, California.

出版信息

Mol Reprod Dev. 1990 Jan;25(1):3-13. doi: 10.1002/mrd.1080250103.

Abstract

A recombinant plasmid containing the Rous sarcoma virus-long terminal repeat (RSV-LTR) promoter linked to rainbow trout (Salmo gairdneri) growth hormone (GH) cDNA was microinjected into fertilized carp eggs. Genomic DNA extracted from pectoral fin of individual presumptive transgenic fish was analyzed by dot blot and Southern blot hybridization, using the RSV-LTR and/or the GH cDNA sequences as probes. Out of 365 presumptive transgenic fish analyzed, 20 individuals were found to contain pRSV-rtGH-cDNA sequence in the genomic DNA. Expression of the trout GH polypeptide was detected by immunobinding assay in the red blood cells of nine transgenic fish tested. The level of expression, however, varied among the transgenics and could not be correlated with exogenous DNA copy number. Although there was considerable variation in the sizes of the transgenic fish, those microinjected during the one-cell stage were (P less than 0.05) 22% larger, on the average, than their sibling controls. A randomly selected fraction of the progeny derived from crosses between transgenic males and non-transgenic females inherited the foreign DNA. These transgenic progeny grew faster (P less than 0.05) than their non-transgenic siblings.

摘要

将一个含有与虹鳟鱼(Salmo gairdneri)生长激素(GH)cDNA相连的劳氏肉瘤病毒长末端重复序列(RSV-LTR)启动子的重组质粒显微注射到鲤鱼受精卵中。以RSV-LTR和/或GH cDNA序列为探针,通过斑点杂交和Southern杂交对从单个推定转基因鱼的胸鳍中提取的基因组DNA进行分析。在分析的365条推定转基因鱼中,发现有20条个体的基因组DNA中含有pRSV-rtGH-cDNA序列。通过免疫结合试验在9条受试转基因鱼的红细胞中检测到了鳟鱼GH多肽的表达。然而,转基因鱼之间的表达水平各不相同,且与外源DNA拷贝数无关。虽然转基因鱼的大小差异很大,但在单细胞阶段显微注射的鱼平均比它们的同胞对照大22%(P<0.05)。从转基因雄性与非转基因雌性杂交后代中随机选取的一部分继承了外源DNA。这些转基因后代比它们的非转基因同胞生长得更快(P<0.05)。

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