吉富罗非鱼两种生长激素受体基因的分离及与增重相关的SNPs位点  

Isolation of Two Growth Hormone Receptor Genes and SNPs Associated with Body Weight in GIFT Strain Tilapia Oreochromis niloticus

作　　者：阮瑞霞[1] 俞菊华[1,2] 李红霞[2] 李建林[2] 唐永凯[2] 魏可鹏[1] 

机构地区：[1]南京农业大学无锡渔业学院,无锡214081 [2]中国水产科学研究院淡水渔业研究中心农业部淡水鱼类遗传育种和养殖生物学重点开放实验室,无锡214081

出　　处：《动物学杂志》2011年第3期37-46,共10页Chinese Journal of Zoology

基　　金：中央级公益性科研院所基本科研业务费专项资金(No.2011JBFC03,2009JBFB05);农业部淡水生物多样性保护与利用重点开放实验室第七期开放课题(No.LFBU0706);公益性行业(农业)科研专项经费项目(No.200903046-02)

摘　　要：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。Growth hormone receptors（GHR） which located on the cell membrane is the only pathway for growth hormone（GH） to get into cells and affect the growth in animals,they play a vital important role on animal growth.Two cDNA sequences and DNA sequences of exon 2-9 of GHR1,exon 2-8 of GHR2 were isolated from GIFT strain（Genetically Improved Farmed Tilapia strain,Oreochromis niloticus） using RT-PCR and PCR in this study.Six and sixteen SNPs were identified on GHR1 and GHR2 respectively,only two of which were located at exons.Genotypes were test in eight 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） of 120 individuals from five families of GIFT strain by PCR-RFLP.Correlation analysis between genotypes and weight gain showed that GHR1 intron 3_A612G and intron 3_A989T associated with male fish weight gain（P0.01 and P0.05） significantly.Similar situation was found in the female population,but the difference was not significant（P0.05）.Intron 7_A599C was significantly associated with both male（P0.01） and female（P0.05） fish weight gain.The CC type fish weight gain was significantly higher than AA type＇s.On the GHR2,only intron 3_G687T was found to correlate with female weight gain significantly（P0.05）.The GT type female fish gain was significantly higher than GG type＇s.The correlation between genotypes of 3_A612G,3_A989T,7_A599C and 3_G687T and weight gain was detected in 356 individuals from 60 families of GIFT strain（6 individuals in average of each family）.There were similar growth situation among different genotypes＇ fish,the decreased significance may result in the diverse genetic background and smaller number of samples.Real-time quantitative PCR results indicated that GHR1 expression in liver of male fish was 1.36 times higher than that in female＇s liver,the male fish GHR2 expression in the tissues（except kidney） was lower than that in the female,which can explain the different eff

关 键 词：吉富罗非鱼 生长激素受体 单核苷酸多态性 增重 组织表达 

分 类 号：S917.4[农业科学—水产科学]