论文部分内容阅读
应用RT-PCR及PCR分离到鲤鱼2个jlGHSR2s基因,jlGHSR2a和2b,分别编码366,367个氨基酸,相似性高达98%,和金鱼、斑马鱼GHSR2的相似性高达95%。jlGHSR2s基因的内含子位于起始密码子793 bp后,2a和2b内含子的长度和序列均存在明显差异。试验首次发现了jlGHSR2a的转录变体(Variant)jlGHSR2a’,由选择性剪切靠近内含子的57 bp外显子1和保留靠近外显子2的28 bp内含子构成,编码350个氨基酸,缺失最后2个跨膜区,剪切序列两端碱基为CT-AC。荧光定量RT-PCR结果表明,jlGHSRs在脑和精巢表达量最高,1s在检测的组织均有表达,但2s在其他组织表达量很少。jlGHSRs的转录变体1s’和2a’的表达量与1s和2s有一定的正相关性,但1s/1s’在各组织不一致,比值最高出现在肝脏,其次是肌肉、肠和脑,精巢和卵巢最低。生长对脑中jlGHS-R1s的表达量没有显著影响,但生长慢的个体肝和肠中jlGHSR1s的表达量比生长快的高,1s/1s’也有明显升高;肌肉中则相反,1s/1s’在雄鱼肌肉中减小明显。饥饿使雌鱼脑、雌鱼前肠和雌雄鱼后肠jlGHSR1s的表达量显著降低,对其余组织中的表达则不显著。生长快慢或饥饿对脑中jlGHSR2s的表达量均没有显著影响。结果表明,jlGHSR1s的表达量与生长有明显的相关性。
Two jlGHSR2s genes, jlGHSR2a and 2b, were isolated from the common carp by RT-PCR and PCR, respectively, encoding 366,367 amino acids with a similarity of 98%. The similarity to goldfish and zebrafish GHSR2 was 95%. The introns of jlGHSR2s gene located at the start codon of 793 bp, 2a and 2b intron length and sequence were significantly different. For the first time, a variant of jlGHSR2a (Variant) jlGHSR2a was found. It consisted of a 57 bp exon 1 that was cleaved closer to the intron and a 28 bp intron close to the exon 2, encoding 350 amino acids , The last two transmembrane regions were deleted. The bases of both ends of the cleavage sequence were CT-AC. Fluorescent quantitative RT-PCR results showed that jlGHSRs had the highest expression level in brain and testis, 1s was detected in all tested tissues, but 2s was less expressed in other tissues. The expression levels of 1s’ and 2a ’in jlGHSRs were positively correlated with 1s and 2s, but 1s / 1s’ was not consistent in all tissues, with the highest ratio in liver, followed by muscle, intestine and brain, testis and The lowest ovary. Growth had no significant effect on the expression of jlGHS-R1s in brain. However, the expression of jlGHSR1s in liver and intestine of slow-growing individuals was higher than that of fast growth, and also significantly increased at 1s / 1s’. In muscle, on the contrary, 1s / 1s Reduced significantly in male muscle. The hunger caused a significant decrease in the expression of jlGHSR1s in the brain, females, foregut and males and females, but not in other tissues. Growth rate or hunger had no significant effect on the expression of jlGHSR2s in brain. The results showed that the expression of jlGHSR1s significantly correlated with the growth.