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短讯:猪5-氨基乙酰丙酸合酶1(ALAS1)基因的分子克隆、表达模式及其与繁殖性状的关联

Short Communication: Molecular cloning and expression pattern of the porcine 5-aminolevulinate synthase 1 (ALAS1) gene and its association with reproductive traits.

作者信息

Liu L Q, Li F E, Deng C Y

机构信息

Institute of Animal Sciences and Veterinary Medicine, Anhui Academy of Agricultural Sciences, Hefei, Anhui, China.

Key Laboratory of Pig Genetics and Breeding of Ministry of Agriculture and Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction of Ministry of Education, Huazhong Agricultural University, Wuhan, Hubei, China.

出版信息

Genet Mol Res. 2016 Feb 22;15(1):gmr7544. doi: 10.4238/gmr.15017544.

Abstract

5-Aminolevulinate synthase 1 (ALAS1) is the first enzyme in the heme biosynthetic pathway and is upregulated in follicular tissue during the early stages of ovulation. In this study, we isolated the complete coding sequence of the porcine ALAS1 gene and its 2-9 intron sequence, identified a single nucleotide polymorphism (SNP; T/C) in intron 9, and developed a PCR-MspI-restriction fragment length polymorphism genotyping assay. Association of the SNP with litter size was assessed in two populations [purebred Large White and the experimental synthetic (DIV) line]. Statistical analysis demonstrated that for total number of piglets born (TNB) in all parities, pigs with the CC genotype had an additional 0.68 and 0.74 piglets compared to the TC and TT animals (P < 0.05) in the DIV line, respectively. Purebred Large White sows inheriting the CC and TC genotypes gave birth to an additional 0.96 and 0.70 piglets compared to the TT animals (P < 0.05) in all parities, respectively. In addition, for TNB in all parities, a significant additive effect of 0.48 ± 0.23 and 0.37 ± 0.17 piglets/ litter was detected in sows of both populations (P < 0.05), respectively. The highest levels of ALAS1 gene expression were observed in isolated ovarian granulosa cells 2 and 12 h after stimulation with pregnant mare serum gonadotropin human chorionic gonadotropin, which represents the time of follicular development and ovulation, respectively. Therefore, the ALAS1 gene was significantly associated with litter size in two populations and could be a useful molecular marker for the selection of increasing litter size in pigs.

摘要

5-氨基酮戊酸合酶1(ALAS1)是血红素生物合成途径中的首个酶,在排卵早期卵泡组织中上调。在本研究中,我们分离出猪ALAS1基因的完整编码序列及其2-9号内含子序列,在9号内含子中鉴定出一个单核苷酸多态性(SNP;T/C),并开发了一种PCR-MspI-限制性片段长度多态性基因分型检测方法。在两个群体[纯种大白猪和实验合成(DIV)系]中评估了该SNP与产仔数的关联。统计分析表明,在DIV系中,对于所有胎次的总产仔数(TNB),CC基因型的猪比TC和TT基因型的猪分别多0.68头和0.74头仔猪(P<0.05)。在所有胎次中,继承CC和TC基因型的纯种大白母猪比TT基因型的母猪分别多产0.96头和0.70头仔猪(P<0.05)。此外,对于所有胎次的TNB,在两个群体的母猪中分别检测到显著的加性效应,即每窝0.48±0.23头和0.37±0.17头仔猪(P<0.05)。在用孕马血清促性腺激素和人绒毛膜促性腺激素刺激后2小时和12小时,在分离的卵巢颗粒细胞中观察到ALAS1基因表达的最高水平,这分别代表卵泡发育和排卵的时间。因此,ALAS1基因在两个群体中与产仔数显著相关,可能是用于选择增加猪产仔数的有用分子标记。

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