The A746G (FecB) mutation in BMPR1B significantly affects the ovulation rate in Booroola-Merino sheep (Mulsant et al.
In this study, we constructed a vector containing the porcine BMPR1B CDS with the mutant allele (G) at the FecB site by site-directed mutagenesis, and then introduced the vector into primary porcine fetal fibroblasts (PFF) by liposome-mediated transfection.
Detection of the BMPR1B A746G mutation in diverse pig breeds
DNA samples of 20 pigs (10 females and 10 males) each from 3 Western breeds (Large White, Landrance, and Duroc), 11 Chinese local breeds (Bama Xiang, Baoshan Big-Ear, Luchuan, Min, Jiaxin Black, Mingguang Small-Ear, Wuzhishan, Laiwu, Jinhua, Tibetan pig and Chinese wild boar) were used to genotype the BMPR1B A 746G mutation.
The synthesized cDNA was used as template to amplify the porcine BMPR1B coding region using the gene-specific primers CDS-F1 and CDS-R1 (Table 1), which were designed based on the mRNA sequence of porcine BMPR1B gene (GenBank accession no.
Introduction of the mutated BMPR1B gene into porcine fetal fibroblasts
The DNA was used to amplify the vector region spanning the promoter and BMPR1B CDS using N2-F and N2-R primers (Table 1).
1] female siblings were slaughtered at 3 days of age to collect tissues for expression analyses of the BMPR1B gene.