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实验分离了吉富罗非鱼(Oreochromis niloticus)两种生长激素受体基因GHR1和GHR2的全长cDNA以及GHR1阅读框中外显子2～9、GHR2阅读框外显子2～8的DNA序列。在GHR1和GHR2分别找到6个和16个SNPs位点,其中只有2个位点在外显子部分。使用PCR-RFLP方法检测了5个家系共120尾吉富罗非鱼在8个SNPs(GHR1内含子3_A612G、内含子3_A989T、内含子7_A599C;GHR2内含子3_C330T、内含子3_A645G、内含子3_G687T、内含子3_A967G、内含子7_C107T)的基因型。不同基因型与雌、雄吉富罗非鱼增重的相关性分析显示,GHR1内含子3_A612G及内含子3_A989T与雄鱼增重极显著(P<0.01)和显著(P<0.05)相关,杂合型个体增重最大,在雌鱼群体中有类似趋势,但差异不显著;内含子7_A599C与雄鱼、雌鱼增重分别呈现极显著(P<0.01)和显著(P<0.05)相关,CC型雌雄鱼增重均大于AA型;GHR2上只有内含子3_G687T与雌鱼增重显著相关(P<0.05),GT型个体增重显著大于GG型。检测了60个家系共356尾吉富罗非鱼在3_A612G、3_A989T、7_A599C和3_G687T的基因型与增重性状的相关性,结果具有相同趋势,因样本少、遗传背景多样,导致差异性显著降低。实时定量PCR结果显示,GHR1在雄鱼肝的表达量是雌鱼的1.36倍;除肾外,GHR2在雄鱼各组织的表达量均低于雌鱼,这部分解释了GHR1和GHR2基因上SNP位点对雌、雄吉富罗非鱼增重的不同影响。此外,GHR1、GHR2对生长影响程度的不同,导致只在GHR2上找到1个SNP位点对增重有影响。
The full length cDNA of two growth hormone receptor genes GHR1 and GHR2 of Oreochromis niloticus and the DNA sequences of exon 2 to amino acid 9 of GHR1 and exons 2 to 8 of GHR2 reading frame in GHR1 reading frame were isolated. Six and 16 SNPs were found in GHR1 and GHR2, respectively, of which only two were in the exon. PCR-RFLP was used to detect a total of 120 SNPs in 5 families with 8 SNPs (GHR1 intron 3_A612G, intron 3_A989T, intron 7_A599C; GHR2 intron 3_C330T, intron 3_A645G, Intron 3_G687T, intron 3_A967G, intron 7_C107T) genotypes. Correlation analysis between different genotypes and weight gain of male and male O. tilapia showed that GHR1 intron 3_A612G and intron 3_A989T were significantly associated with male weight gain (P <0.01) and significantly (P <0.05) Heterozygous individuals had the highest weight gain and similar trend in females, but the difference was insignificant. Intron 7_A599C and male and female weight gain showed significant (P <0.01) and significant (P <0.05) (P <0.05). Only the intron 3_G687T of GHR2 was significantly associated with the weight gain of the female (P <0.05). The weight gain of the GT genotype was significantly greater than that of the GG type. The genotypes and weight gain traits of 356 Aphids tilapia from 60 families were detected in 3_A612G, 3_A989T, 7_A599C and 3_G687T. The results showed the same trend, and the differences were notable due to the small sample size and diverse genetic backgrounds. Real-time quantitative PCR results showed that the expression level of GHR1 in male livers was 1.36 times of that of females; except for the kidney, the expression of GHR2 in males was lower than that in females, which partly explained that SNPs in GHR1 and GHR2 genes Different Effects of Site on Weight Gain of Female and Male. In addition, GHR1, GHR2 on the growth of different degrees, resulting in only one found in GHR2 SNP sites have an impact on weight gain.