畜牧兽医学报 ›› 2010, Vol. 41 ›› Issue (9): 1082-1089.doi:

• 遗传繁育 • 上一篇    下一篇

鸭BLVRA基因cDNA克隆及其在输卵管子宫部的mRNA表达量与青壳性状的相关性分析

张宝乐1,2, 李国勤1,章鹤1,2,沈军达1,李进军1,陶争荣1,陈奕春3,
王德前1,陈文标4,石放雄2*,卢立志1*
  

  1. 1.浙江省农业科学院畜牧兽医研究所,杭州 310021;2.南京农业大学动物科技学院,南京 210095;3.缙云县畜牧兽医局,缙云 321403;4.浙江省五莲农牧有限公司,缙云321403
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2010-09-20 发布日期:2010-09-20
  • 通讯作者: 卢立志,石放雄

between mRNA Expression of Duck BLVRA Gene in Interstitial Part of Uterine Tube and Green Shell Character

ZHANG Bao-le1,2, LI Guo-qin1, ZHANG He1,2 , SHEN Jun-da1, LI Jin-jun1, TAO Zheng-rong1, CHEN Yi-chun3, WANG De-qian1, CHEN Wen-biao4, SHI Fang-xiong2*, LU Li-zhi1*   

  1. 1. Institute of Animal Husbandry and Veterinary Medicine, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China; 2. College of Animal Science and Technology, Nanjing Agricultural University, Nanjing 210095, China; 3. Jinyun Husbandry and Veterinary Bureau, Jinyun 321403, China; 4. Zhejiang Wulian Agriculture and Animal Co., Ltd,Jinyun 321403, China
  • Received:1900-01-01 Revised:1900-01-01 Online:2010-09-20 Published:2010-09-20

摘要: 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1 071个核苷酸组成,编码302个氨基酸,等电点为7.15,分子量为34.3 ku,分别将已有编码区序列向5′和3′端延伸238和199 bp,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。

Abstract: 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 5′ ends 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 BLVRA were 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 BLVRA was comprised of 1 071 nucleotides, encoding 302 amino acids and having the isoelectric point of 7.15 and the molecular weight of 34.3 ku. The 5′ and 3′ ends of the duck BLVRA gene sequence published in the GenBank were extended 238 and 199 bp, respectively. Comparing the putative amino acid of duck BLVRA gene 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 BLVRA gene was conservative in the process of evolution. The relative gene expression level of BLVRA in white eggshells of Jinyun duck was 3 times 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 BLVRA gene relative expression levels(r=0.719,P<0.05). Together, the result suggested that BLVRA gene could be used as a candidate gene for molecule breeding of the green shell duck.