Anas platyrhynchos BLVRA Gene:Molecular Cloning and Correlation between Green Shell Character and mRNA Expression in the Shell Gland in the Duck

This study has examined the structure and function of the biliverdin reductase A (BLVRA) gene and its correlation with the green eggshell character in Jinyun ducks. The previously unknown sequences of 3' and 5' ends of duck BLVRA mRNA were cloned, using RT-PCR and 5' RACE methods with four pairs of primers designed from the known coding sequence (CDS) of the duck BLVRA gene. Gene expression levels of BLVRA in the shell gland were quantified by real-time PCR, and eggshell colors were determined objectively by the reflection coefficient. The cDNA sequence of BLVRA was comprised of 1071 nucleotides, encoding a protein of 303 amino acids with an isoelectric point of 7.15 and molecular weight of 34.3 kDa. The 5' and 3' ends of the duck BLVRA gene sequence published in the GenBank were extended by 238 and 199 bp, respectively. The deduced amino acid sequence of duck BLVRA had 95.3% identity with Gallus gallus, 95.0% with Taeniopygia guttata, 70.0% with Xenopus tropicalis,61.1% with Bos taurus, 60.5% with Homo sapiens and 59.5% with Mus musculus, which indicated that the BLVRA gene was evolutionarily conserved. The relative gene expression level of BLVRA in shell glands of Jinyun ducks with white eggshells was 3 times higher than that in ducks with dark green eggshells (P<0.01). There was a significant positive correlation between reflection coefficient of eggshells and the relative expression of the BLVRA gene (r=0.719, P<0.05). Together, these findings suggest that the BLVRA gene could be a candidate gene for the molecule breeding of ducks with green eggshells.

新生儿不明原因高胆红素血症BLVRA基因突变分析

目的探讨分析新生儿不明原因高胆红素血症胆绿素还原酶A(biliverdin reductase A,BLVRA)基因突变情况。方法选取2021年7月至2023年7月我院出生并诊断为不明原高胆红素血症新生儿52例为观察组,并选取同期50例健康新生儿作为对照组。新生儿分娩3d后,取其血清样本检测BLVRA基因突变情况,分析BLVRA基因rs699512位点突变与新生儿不明原因高胆红素血症的关系。结果观察组新生儿分娩3d后总胆红素水平为298.65±45.72μmol/L;对照组总胆红素水平为166.82±39.57μmol/L;观察组新生儿分娩3d后总胆红素水平高于对照组(P<0.05)。观察组BLVRA基因rs699512位点AA基因型、等位基因A频率分布高于对照组(P<0.05)。AA基因型新生儿血清总胆红素水平低于AG基因型和GG基因型患儿,且组间比较差异具有统计学意义(P<0.05);并且,观察组不同基因型患儿分娩3d后血清总胆红素水平均高于对照组(P<0.05)。AA基因型总胆红素水平≥332μmol/L的发生率高于AG基因型和GG基因型(P<0.05)。结论BLVRA基因突变可引起新生儿不明原因高胆红素血症患儿血清总胆红素水平升高,rs699512位点AA基因型多态性可能参与新生儿不明原因高胆红素血症发病过程。

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

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。

麻旺鸭的胆绿素还原酶基因SNP发现及表达分析

为了研究胆绿素还原酶A(BLVRA)基因在代谢过程中与蛋壳颜色的关系,研究对13只麻旺鸭的输卵管部BLVRA基因进行了克隆测序,并检测该基因在不同组织及不同个体的子宫部的表达量。结果表明:共发现BLVRA基因2个SNP位点;BLVRA基因在卵巢、肺脏、腿肌、脾脏、肝脏、心脏、肾脏、胸肌、输卵管9种组织中均有表达,其中卵巢、肺脏、腿肌、胸肌中表达量高于其他组织。