title: Polymorphism analysis of BMPR1B gene by forced RFLP and PCR-SSCP techniques and expression of the mutation in introgressed sheep.
abstract: The present study was conducted to screen Kashmir valley sheep with history of prolificacy for the presence of FecB mutation. Forced polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and single strand conformation polymorphism (SSCP) techniques were employed to detect any polymorphism present in bone morphogenetic protein receptor type 1B (BMPR1B) gene. Further, it was aimed at introgressing the FecB mutation into nonprolific noncarrier sheep. A 140-bp fragment of BMPR1B gene was amplified from isolated genomic DNA and subjected to forced RFLP with restriction enzyme AvaII. Three different RFLP patterns were identified. SSCP analysis showed one-to-one correspondence with RFLP patterns. Sequencing of the samples showing different patterns revealed that the wild (+) and mutant (B) alleles were different by a single nucleotide substitution in the form of A109G from wild to mutant allele. It led to change in amino acid from Glutamine (Q) to Arginine (R) from wild to mutant allele. The mutation was only detected in NARI-Suwarna and their crosses; all Kashmir valley sheep with prolific history lacked it. The + allele was abundant in the studied population. The FecB mutation was introgressed in nonprolific noncarrier sheep by crossing ewes with NARI-Suwarna rams possessing the mutation. First generation crossing produced heterozygous (B+) progeny. Some of the F(1) heterozygous ewes gave birth to twins when mated to unrelated NARI-Suwarna rams. It showed that FecB mutation was successfully expressing in those crosses.
authors.affiliation: Division of Animal Genetics and Breeding, Faculty of Veterinary Sciences & Animal Husbandry, Sher-e-Kashmir University of Agricultural Sciences & Technology of Kashmir, Shuhama, Alusteng, Srinagar, Jammu & Kashmir, India.
authors.forename: Tasleem Ahmad Shamas
authors.initials: TA
authors.lastname: Ganai
authors.forename: Siddhartha Sarathi
authors.initials: SS
authors.lastname: Misra
authors.forename: Mir
authors.initials: M
authors.lastname: Shabir
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D052004:Bone Morphogenetic Protein Receptors / Q000235:genetics*; D004247:DNA / Q000737:chemistry / Q000235:genetics; D005298:Fertility / Q000235:genetics*; D014644:Genetic Variation; D005838:Genotype; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000662:veterinary; D012150:Polymorphism, Restriction Fragment Length; D020641:Polymorphism, Single Nucleotide; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D012756:Sheep / Q000235:genetics*
chemical_list: D004247:DNA; D052004:Bone Morphogenetic Protein Receptors
references.citation: Anim Reprod Sci. 2008 May;105(3-4):319-31
references.pmid: 17449205
references.citation: Biol Reprod. 2001 Apr;64(4):1225-35
references.pmid: 11259271
references.citation: Anal Biochem. 1991 Jul;196(1):80-3
references.pmid: 1716076
references.citation: Anim Reprod Sci. 2007 May;99(1-2):44-52
references.pmid: 16859845
references.citation: Anim Reprod Sci. 2006 Mar;92(1-2):87-96
references.pmid: 15982834
references.citation: Reprod Fertil Dev. 2006;18(4):433-7
references.pmid: 16737636
references.citation: Trop Anim Health Prod. 2010 Jun;42(5):985-93
references.pmid: 20020203
references.citation: Anim Reprod Sci. 2009 Nov;116(1-2):1-9
references.pmid: 19201555