Abstract: In order to clarify the basic properties of leptin gene in river puffer Takifuguobscurus, and to explore the polymorphism of the leptin gene and its correlation with the growth of the puffer, the cDNA sequence of the leptin gene was cloned in 6 samples of 3 month old puffer by RT-PCR. Then the SNP loci related to growth in the leptin gene were screened in 178 samples of 8 month old river puffer by direct sequencing method. The results showed that the cloned leptin cDNA sequence was 661 bp in length, including an open reading frame of 459 bp, encoding a total of 152 amino acids (hydrophilic stable protein), containing a signal peptide sequence of 19 amino acids. There was high tertiary structural similarity (96.7%) between river puffer T.obscurus and other fish species, with the maximal similarity of Leptin amino acid sequence between river puffer T.obscurus and tiger puffer Takifugurubripes and weak similarity of Leptin amino acid sequence between puffer T.obscurus and other species (including grass carp Ctenopharyngodonidella 25.25%). Two SNP loci with stable typing (T24G, T193A) were screened on the leptin gene of river puffer T.obscurus, in which there was significantly different body weight in the individuals with different genotypes at the T24G locus (P<0.05), significantly smaller in GG genotype than that in TG and TT genotype. The association analysis of combination of the different genotypes of the two SNP loci revealed that the dominant double-type D5 individuals had the maximal average value of the three growth traits, and significantly higher body mass than that in D2 and D4 individuals (P<0.05), very significantly higher than that in D6 individuals (P<0.01). There was significantly higher body length and total length in D5 individuals than that in D6 individuals (P<0.05). The findings indicate that TG and TT genotype in T24G locus and double-type D5 can be used as candidate molecular markers for assisted breeding of river puffer T.obscurus.
余云登, 朱浩拥, 王盛南, 王耀辉, 马文超, 杨晓, 朱永祥, 钱晓明, 王秀利. 暗纹东方鲀leptin基因的克隆、SNP筛选及其与生长性状的关联分析[J]. 大连海洋大学学报, 2021, 36(2): 207-213.
YU Yundeng, ZHU Haoyong, WANG Shengnan, WANG Yaohui, MA Wenchao, YANG Xiao, ZHU Yongxiang, QIAN Xiaoming, WANG Xiuli?. Cloning, SNP screening and association with growth traits of leptin gene in river puffer Takifugu obscurus. Journal of Dalian Ocean University, 2021, 36(2): 207-213.