Polymorphism analysis of BMPR1B gene by forced RFLP and PCR-SSCP techniques and expression of the mutation in introgressed sheep |
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| Authors: | Ganai Tasleem Ahmad Shamas Misra Siddhartha Sarathi Shabir Mir |
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| Institution: | (1) 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, 190006, Jammu & Kashmir, India; |
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| 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 F1 heterozygous ewes gave birth to twins when mated to unrelated NARI-Suwarna rams. It showed that FecB mutation was successfully expressing in those crosses. |
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