Journal of Stress Physiology & Biochemistry, Vol. 16 No. 2 2020, pp. 81-85    ISSN 1997-0838
Original Text Copyright (cc) 2020 by  Sachin, Behnaz, Anjali and Anushree



ORIGINAL ARTICLE
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PCR Based Detection of Insulin Like Protein from Dolichos lablab L.

Palekar Sachin 1*, Patel Behnaz2, Apte-Deshpande Anjali3 and Lokur Anushree4

1 Dept of Bioanalytical Sciences, Ramnarain Ruia Autonomous College, Mumbai, India
2 Dept of Botany, Ramnarain Ruia Autonomous College, Mumbai, India
3 Central Dogma pvt. Ltd, Pune, India
4 Department of Microbiology, Ramnarain Ruia Autonomous College, Mumbai, India

*E-Mail: sachinpalekar@ruiacollege.edu


Received February 29, 2020


In the current research work, the genetic region corresponding to both alpha and beta chains of insulin like protein from Dolichos lablab L was identified using PCR. Earlier study by Venāncio et al., (2003) has revealed that the amino acid sequence of alpha and beta chains of bovine insulin has significant similarity to amino acid sequence of insulin like protein from legume Vigna unguiculata (cowpea). Although the amino acid sequence was available for V. unguiculata, the corresponding DNA sequence was not available in the database. Considering the resemblance in protein sequence, primers were designed using nucleotide sequences of alpha and beta chains of bovine insulin and were used for amplification of these chains from V. unguiculata and D. lablab. Genomic DNA extraction was carried out by the conventional ethanol precipitation method followed by purification using preparative AGE. Standardization on PCR amplification of alpha and Beta chain of Insulin from D. lablab L. and V. unguiculata was carried out. Results obtained for the PCR amplification of alpha and beta chains were validated by the re-amplification of gene, where the previously obtained PCR product was used as template DNA. All the above experiments suggest that the DNA sequence of both chains of insulin gene is probably highly conserved amongst different organisms as the primer designed using bovine insulin DNA sequence could amplify the same gene in plants like V. unguiculata and D. lablab L.

Key words:    Insulin like protein, Bean, PCR based detection, Dolichos lablab

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