Chen T T, Kight K, Lin C M, Powers D A, Hayat M, Chatakondi N, Ramboux A C, Duncan P L, Dunham R A
Department of Biological Sciences, Univesity of Maryland, Baltimore 21202.
Mol Mar Biol Biotechnol. 1993 Mar-Apr;2(2):88-95.
Transgenic common carp, Cyprinus carpio, possessing the long terminal repeat (LTR) sequence of avian Rous sarcoma virus (RSV) fused to the rainbow trout (rt) growth hormone (GH1) complementary DNA (cDNA) were produced by microinjection. Initial studies showed that the transgenic common carp transmitted the foreign DNA to a significant fraction of their progeny in three of four crosses of transgenic males with control females. These progeny grew 20 to 40% faster than their nontransgenic full siblings. In this study, additional experiments were conducted to evaluate inheritance and expression of the foreign GH gene in transgenic common carp, and the growth performance of these transgenic fish. Four P1 (parental generation produced by microinjection) x nontransgenic controls, four P1 x P1, and one P1 x F1 matings of transgenic carp containing RSVLTR-rtGH1 cDNA were made. The percentages of transgenic progeny resulting from these matings were: 0, 32, 42, 100 (4 progeny only), 21, 21, 31, 30, and 23%, respectively. All crosses except 1 siblot (control x P1) exhibited progeny ratios below the expected 50 or 75% transgenic. These results indicate that most of these transgenic P1 had the foreign gene in their germ line but were mosaics, and at least one transgenic individual did not have the RSVLTR-rtGH1 cDNA in the gonadal tissue. Both P1 and F1 transgenic fish produce trout growth hormone mRNA and polypeptide as determined by reverse transcription polymerase chain reaction amplification, RNA dot-blot hybridization, and radio-immunobinding assay. Growth response by families of F1 transgenic fish to the addition of rtGH1 cDNA varied widely.(ABSTRACT TRUNCATED AT 250 WORDS)
通过显微注射产生了转基因鲤鱼(Cyprinus carpio),其具有与虹鳟鱼(rt)生长激素(GH1)互补DNA(cDNA)融合的禽劳斯肉瘤病毒(RSV)的长末端重复序列(LTR)。初步研究表明,在转基因雄性与对照雌性的四次杂交中的三次杂交中,转基因鲤鱼将外源DNA传递给了相当一部分后代。这些后代比它们的非转基因全同胞生长快20%至40%。在本研究中,进行了额外的实验来评估转基因鲤鱼中外源生长激素基因的遗传和表达,以及这些转基因鱼的生长性能。对含有RSVLTR-rtGH1 cDNA的转基因鲤鱼进行了四次P1(显微注射产生的亲代)×非转基因对照、四次P1×P1以及一次P1×F1交配。这些交配产生的转基因后代的百分比分别为:0、32、42、100%(仅4个后代)、21、21、31、30和23%。除了1个同胞杂交(对照×P1)外,所有杂交的后代比例均低于预期的50%或75%转基因比例。这些结果表明,这些转基因P1中的大多数在其生殖系中含有外源基因,但为嵌合体,并且至少有一个转基因个体在性腺组织中不含有RSVLTR-rtGH1 cDNA。通过逆转录聚合酶链反应扩增、RNA斑点杂交和放射免疫结合测定确定,P1和F1转基因鱼均产生鳟鱼生长激素mRNA和多肽。F1转基因鱼家族对添加rtGH1 cDNA的生长反应差异很大。(摘要截短于250字)