Abstract
Insulin plays an important role in maintaining the blood glucose level of the body. The β-cells of pancreas produce insulin in the form of precursor that is preproinsulin. The gene of preproinsulin provides an interesting system for addressing question related to molecular evolution. Recombinant DNA technology has made it possible to isolate and sequence the chromosomal genes coding for unique protein products. Although preproinsulin of various organism has been isolated and cloned, but there is no report from buffalo (Bubalus bubalis) that is our major livestock. The genomic DNA of buffalo was isolated using Laura-Lee-Boodram method. The part of preproinsulin gene (596bp and 520bp) using BPPI-UPS and bpiful_F as forward and BC1-C as reverse primer was amplified. Cloning of amplified fragments of gene were performed in pCR 2.1 vector. Positive clones were screened on the basis of blue white selection. The band obtained on 596bp and 520bp after colony PCR confirmed the successful cloning of preproinsulin gene in pCR 2.1 vector.
Highlights
Diabetes mellitus is a metabolic disease in which blood glucose level of the person is high because the body does not produce insulin or the cells do not respond to the insulin which is produced in the body [1]
The band obtained on 596bp and 520bp after colony PCR confirmed the successful cloning of preproinsulin gene in pCR 2.1 vector
In eukaryotes insulin is synthesized in the beta cells of pancreas in the form of its precursor, preproinsulin
Summary
Diabetes mellitus is a metabolic disease in which blood glucose level of the person is high because the body does not produce insulin or the cells do not respond to the insulin which is produced in the body [1]. High blood glucose level stimulates the β- cells of pancreas to release insulin, which allows the glucose to enter the body’s cell and it is used as a fuel for energy. It maintains the blood sugar level in the range of 80-120mg/dl. Beta cells release insulin in response to change in glucose level. The increase in calcium ions trigger the export of insulin storing granules by exocytosis and this results in the transport of insulin out of the cells in the bloodstream[5]. To fill the gap preproinsulin from buffalo (Bubalus bubalis)
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