Nucleotide Sequencing and PCR-RFLP of Insulin-like Growth Factor Binding Protein-3 Gene in Riverine Buffalo (Bubalus bubalis)

Padma, B.; Kumar, Pushpendra; Choudhary, V.; Dhara, S. K.; Mishra, A.; Bhattacharya, T. K.; Bhushan, B.; Sharma, Arjava

doi:2004.17.7.910

Article

Animal Breeding and Genetics

Asian-Australasian Journal of Animal Sciences 2004;17(7): 910-913.
https://doi.org/10.5713/ajas.2004.910 Published online January 1, 2004.

Nucleotide Sequencing and PCR-RFLP of Insulin-like Growth Factor Binding Protein-3 Gene in Riverine Buffalo (Bubalus bubalis)

B. Padma, Pushpendra Kumar, V. Choudhary, S. K. Dhara, A. Mishra, T. K. Bhattacharya, B. Bhushan, Arjava Sharma

Abstract

Insulin-like growth factor binding protein-3 (IGFBP-3) gene is a structural gene associated with the growth and development of the animals. The present investigation was carried out to unravel nucleotide sequence and polymerase chain reaction-restriction fragment polymorphism (PCR-RFLP) of IGFBP-3 gene in buffalo. Genomic DNA was isolated from a total of 157 animals belonging to Murrah, Surti, Jaffarabadi and Nagpuri breeds of Indian riverine buffalo. A 655 bp of IGFBP-3 gene was amplified in all the breeds and amplicons were digested with Hae III, Taq I and Msp I restriction enzymes. On digestion with Hae III yielded single restriction pattern of 8 fragments of sizes 201, 165, 154, 56, 36, 19, 16 and 8 bp in all the animals studied. Similarly Taq I and Msp I also revealed single restriction pattern yielding fragments of sizes 240 and 415 bp and 145 and 510 bp, respectively. This shows non-polymorphic nature of restriction sites in buffalo. Nucleotide sequencing of 587 bp of IGFBP-3 gene in Murrah buffalo was done and submitted to the GenBank (Accession No. AY304829). Nucleotide sequencing revealed an addition of 4 bases in the intronic region as compared to cattle.

Keywords: Buffalo; IGFBP-3; Sequencing; PCR-RFLP; Polymorphism