Xia Zheng-Long, Yu Ju-Hua, Li Hong-Xia, Li Jian-Lin, Tang Yong-Kai, Ren Hong-Tao, Zhu Shuang-Ning
Nanjing Agricultural University, Wuxi, China.
Yi Chuan. 2013 May;35(5):628-36. doi: 10.3724/sp.j.1005.2013.00628.
Two replicate intestine fatty acid binding protein genes (jlFABP2a and jlFABP2b) were cloned from Cyprinus carpio var. jian using PCR. Both ORFs were 399 bp in length sharing 92.2% similarity with each other, and 88.0% and 90.5% with their counterpart in zebrafish, respectively. The gene structure of jlFABP2s was same as other FABPs, which contained four exons and three introns. Sequences and lengths of introns between 2a and 2b. were obviously different Phylogenetic tree displayed that two jlFABP2s corresponded to one zebrafish FABP2 which matches the fact that the chromosome number of common carp was twice of zebrafish. Real time-PCR showed that jlFABP2 genes mainly expressed in intestine and the expression level was very significantly higher than other tissues such as brain, liver, muscle, kidney, and gonad (P<0.01). The expression level of jlFABP2a was significantly (male, P<0.05) or very significantly (females, P<0.01) higher than 2b in intestine; and 2b was expressed slightly higher than 2a in other tissues. It seemed that 2a expressed specifically in intestine, while 2b expressed ubiquitously. Twelve and four SNP loci were found at jlFABP2a and 2b introns through comparison sequences from 8 individuals, respectively. Genotypes of I1-A15G, I1-A99G, I2-C487T, and I3-A27T on jlFABP2a were detected using PCR-RFLP in selection population of C. carpio var. jian. The SNP genotypes and individual weight gain correlation indicated that four SNPs were significantly (P<0.05 or P<0.01) associated with adult weight gain. Diplotype analysis displayed that individuals with genotype AGGGCCXX or AGGGXXAT grew faster than other individuals by 15%. The individuals with these two genotypes only occupied 9% in total selection populations, indicating the presence of large selection space. The 4 SNPs detected in this experiment can be used in C. carpio var. Jian growth selection breeding plan.
利用聚合酶链式反应（PCR）从建鲤（Cyprinus carpio var. jian）中克隆出两个重复的肠脂肪酸结合蛋白基因（jlFABP2a和jlFABP2b）。两个开放阅读框（ORF）长度均为399 bp，彼此间相似度为92.2%，与斑马鱼对应基因的相似度分别为88.0%和90.5%。jlFABP2s的基因结构与其他脂肪酸结合蛋白（FABPs）相同，包含四个外显子和三个内含子。2a和2b之间内含子的序列和长度明显不同。系统发育树显示，两个jlFABP2s对应于一个斑马鱼FABP2，这与鲤鱼染色体数目是斑马鱼两倍的事实相符。实时定量PCR显示，jlFABP2基因主要在肠道表达，其表达水平显著高于脑、肝、肌肉、肾和性腺等其他组织（P<0.01）。在肠道中，jlFABP2a的表达水平显著高于2b（雄性，P<0.05；雌性，P<0.01）；在其他组织中，2b的表达略高于2a。似乎2a在肠道中特异性表达，而2b广泛表达。通过比较8个个体的序列，在jlFABP2a和2b的内含子中分别发现了12个和4个单核苷酸多态性（SNP）位点。在建鲤选育群体中，利用聚合酶链式反应-限制性片段长度多态性（PCR-RFLP）检测了jlFABP2a上的I1-A15G、I1-A99G、I2-C487T和I3-A27T基因型。SNP基因型与个体体重增加的相关性表明，4个SNP与成年体重增加显著相关（P<0.05或P<0.01）。双倍型分析显示，基因型为AGGGCCXX或AGGGXXAT的个体比其他个体生长速度快15%。这两种基因型的个体在总选育群体中仅占9%，表明存在较大的选择空间。本实验中检测到的4个SNP可用于建鲤生长选育育种计划。
