摘要
为研究胆绿素还原酶A(BLVRA)基因的结构功能,探索其与鸭青壳性状的相关性,根据鸭BLVRA基因部分已知编码序列(CDS),采用RT-PCR和5′RACE方法扩增缙云麻鸭BLVRA基因mRNA的3′和5′末端未知序列,应用荧光定量PCR方法测定鸭输孵管子宫部BLVRA基因表达量,用反射系数法测定蛋壳颜色。结果表明:鸭BLVRA基因cDNA由1071个核苷酸组成,编码302个氨基酸,等电点为7.15,分子量为34.3ku,分别将已有编码区序列向5′和3′端延伸238和199bp,与鸡、珍珠鸟、蟾蜍、牛、人和小鼠的相应氨基酸序列的相似性分别为95.3%、95.0%、70.0%、61.1%、60.5%和59.5%。白壳蛋缙云麻鸭输孵管子宫部BLVRA基因的相对表达量约是深青壳蛋鸭相对表达量的3倍(P<0.01),蛋壳颜色的反射系数与BLVRA基因mRNA相对表达量呈显著的正相关(r=0.719,P<0.05)。初步推断BLVRA基因可作为青壳蛋鸭分子选育的候选基因。
In order to study the structure and function of biliverdin reductase A（BLVRA）gene and its correlation with the character of green shell,the unknown sequences of 3′and 5nds of duck BLVRA mRNA were cloned by using RT-PCR and 5′RACE methods and four pairs of primers were designed and synthesized according to the known coding sequence（CDS）of duck BLVRA gene in the current project.Gene expression levels of BLVRAwere measured by real-time quantitative PCR,and the eggshell colors were determined by the reflection coefficient method.The results showed that the cDNA sequence of BLVRAwas comprised of 1 071nucleotides,encoding 302amino acids and having the isoelectric point of 7.15and the molecular weight of 34.3ku.The 5′and 3nds of the duck BLVRAgene sequence published in the GenBank were extended 238 and 199bp,respectively.Comparing the putative amino acid of duck BLVRAgene with that of other animals showed that it shared 95.3%,95.0%,70.0%,61.1%,60.5%and 59.5%homology with Gallus gallus,Taeniopygia guttata,Xenopus tropicalis,Bos taurus,Homo sapiens and Mus musculus,respectively,which proved that BLVRAgene was conservative in the process of evolution.The relative gene expression level of BLVRAin white eggshells of Jinyun duck was 3times higher than that in dark green eggshells after normalization with GAPDH gene（P 〈0.01）.There was a significant positive correlationship between reflection coefficient of eggshell and BLVRAgene relative expression levels（r=0.719,P〈0.05）.Together,the result suggested that BLVRAgene could be used as a candidate gene for molecule breeding of the green shell duck.
出处
《畜牧兽医学报》
CAS
CSCD
北大核心
2010年第9期1082-1089,共8页
ACTA VETERINARIA ET ZOOTECHNICA SINICA
基金
现代农业产业技术体系建设专项资金
国家"十一五"科技支撑计划课题(2006BAD14B06)
浙江省重大科技攻关项目(2005C12005-01)
