Nucleotide sequencing and PCR-RFLP of insulin-like growth factor binding protein-3 gene in riverine buffalo (Bubalus bubalis)

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. (Asian-Aust. J. Anim. Sci. 2004. Vol 17, No. 7 : 910-913)

胰岛素样生长因子结合蛋白 - 3（IGFBP - 3）基因是一种与动物生长发育相关的结构基因。本研究旨在揭示水牛IGFBP - 3基因的核苷酸序列以及聚合酶链反应 - 限制性片段多态性（PCR - RFLP）。从印度河流型水牛的摩拉、苏尔蒂、贾法拉巴迪和那格浦里品种共157头动物中提取基因组DNA。在所有品种中均扩增出655bp的IGFBP - 3基因，并使用Hae III、Taq I和Msp I限制性内切酶对扩增产物进行酶切。用Hae III酶切时，在所研究的所有动物中均产生8个片段的单一酶切图谱，片段大小分别为201、165、154、56、36、19、16和8bp。同样，Taq I和Msp I也显示出单一的酶切图谱，分别产生大小为240和415bp以及145和510bp的片段。这表明水牛的限制性酶切位点具有非多态性。对摩拉水牛587bp的IGFBP - 3基因进行了核苷酸测序，并提交给GenBank（登录号AY304829）。核苷酸测序显示，与牛相比，其内含子区域增加了4个碱基。（《亚洲 - 澳大利亚动物科学杂志》2004年，第17卷，第7期：910 - 913页）