Sun Caixia, Xie Shuyu, Huang Tao, Zhang Wei, Wang Ansi, Wang Dan, Li Ming, Sun Guirong
College of Animal Science and Veterinary Medicine, Henan Agricultural University, Zhengzhou, Henan 450002, People's Republic of China.
J Genet. 2017 Jun;96(2):313-318. doi: 10.1007/s12041-017-0766-y.
Growth differentiation factor 9 (GDF9) has been shown to be involved in regulating follicular development and reproduction in many mammalian species. However, related information about the effect of the GDF9 gene on reproductive traits of New Zealand white rabbits was rarely reported. In this study, rabbits were distributed into two groups (poor and prolific offspring productions) and cloning and quantitative real-time PCR (qPCR) were employed to characterize the rabbit GDF9 gene. By cloning, 2515-bp genomic DNA and 1359-bp cDNA sequences were obtained. Comparing the two cDNA sequences, three potential mutation sites (C.539C>T,C.562G>C and C.718C>G) in exon 2 of the GDF9 gene were found, and the corresponding amino acids changed (P.183T>M, P.188E>Q and P.240L>V). The qPCR results revealed that GDF9 was not tissue-specific, but rather expressed in all collected tissues. The expression level of the GDF9 gene was highest in the ovary, and was significantly increased (P< 0.05) compared with the other tissues. The liver had the second highest expression, and the heart and spleen had the least expression in New Zealand white rabbits. In the prolific group, the expression quantity of the GDF9 gene significantly increased (P < 0.05) in the heart, spleen, ovary, liver and uterus (P < 0.01) than the other groups. The amino acid sequence identities of human, sheep, goat, mouse, cattle, pig, cat, donkey, Nancy Ma's night monkey and olive baboon were 72, 68, 69, 66, 69, 71, 67, 73, 75 and 73%, respectively. Bioinformatics analysis was executed, and a random coil was determined to be the primary secondary structure.
生长分化因子9（GDF9）已被证明在许多哺乳动物物种中参与调节卵泡发育和繁殖。然而，关于GDF9基因对新西兰白兔繁殖性状影响的相关信息鲜有报道。在本研究中，将兔子分为两组（后代生产性能差和繁殖力高），采用克隆和定量实时PCR（qPCR）技术对兔GDF9基因进行表征。通过克隆，获得了2515 bp的基因组DNA序列和1359 bp的cDNA序列。比较这两个cDNA序列，发现GDF9基因外显子2中有三个潜在突变位点（C.539C>T、C.562G>C和C.718C>G），相应的氨基酸发生了变化（P.183T>M、P.188E>Q和P.240L>V）。qPCR结果显示，GDF9并非组织特异性表达，而是在所有采集的组织中均有表达。GDF9基因的表达水平在卵巢中最高，与其他组织相比显著升高（P<0.05）。在新西兰白兔中，肝脏的表达水平次之，心脏和脾脏的表达水平最低。在繁殖力高的组中，GDF9基因在心脏、脾脏、卵巢、肝脏和子宫中的表达量比其他组显著增加（P<0.05），在子宫中显著增加（P<0.01）。人、绵羊、山羊、小鼠、牛、猪、猫、驴、南希·马氏夜猴和东非狒狒的氨基酸序列同一性分别为72%、68%、69%、66%、69%、71%、67%、73%、75%和73%。进行了生物信息学分析，确定无规卷曲为主要二级结构。
