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Optimization of an efficient SDS-PAGE protocol for rapid protein analysis of Brassica rapa

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Research Paper 01/08/2016
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Optimization of an efficient SDS-PAGE protocol for rapid protein analysis of Brassica rapa

Sohail Ahmad Jan, Zabta Khan Shinwari, Malik Ashiq Rabbani, Sabir Hussain Shah, Muhamad Ishaq Ibrahim, Muhammad Ilyas
J. Bio. Env. Sci.9( 2), 17-24, August 2016.
Certificate: JBES 2016 [Generate Certificate]
Key: Brassica rapaGenetic variationPhylogenetic relationshipSDS-PAGE

Abstract

SDS-PAGE plays a key role in the study of protein based variation among different brassica species. The present study aimed to develop an efficient Sodium Dodecyl Sulphate Poly Acrylamide Gel Electrophoresis (SDS-PAGE) protocol for B. rapa. Ten diverse genotypes were used to study their electrophoretic protein profiling. A thoroughly precise protocol was developed for optimizing the conditions such as proper pH level, centrifugation time, sample size, ammonium per sulphate (APS) concentrations, staining and de-staining time period etc. By optimizing these factors maximum polymorphic proteins were recorded that sizes range from about 10-180 k Da. All the genotypes were classified into four major groups on the basis of similarity that exist. The similarity coefficient value ranges from 40 to 95.2%. The least (40%) and maximum (95.2%) similarity coefficient values were noted among Br-508/Br-728 and Br-695/Br-725 respectively. A robust and quick SDS-PAGE protocol was developed; it will be used to study genetic diversity of other crop species and to widen the agriculture breeding program.

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Akbar F, Yousaf N,  Rabbani MA, Shinwari ZK,  Masood S. 2012. Study of total Seed Proteins Pattern of Sesame (Sesamum indicum L.) Landraces via Sodium Dodecyl Sulfate Polyacrylamide Gel Elecrophoresis (SDS-PAGE). Pakistan Journal of Botany 44(6), 2009-2014.

Arif M, Khurshid H, Siddiqui SU, Jatoi SA, Jan SA, Ilyas M, Khan SA, Khan A, Ibrahim MI, Saleem N, Ghafoor A. 2015. Estimating spatial population structure through quantification of oil content and phenotypic diversity in Pakistani Castor Bean (Ricinus communis L.) germplasm. Science, Technology and Development 34(3), 147-154. http://dx.doi.org/10.3923/std.2015.147.154

Das S, Mukherjee KK. 1995. Comparative study on seed protein of Ipomoea. Seed Science and Technology 23, 501-509.

Dhawale, RN, Mahalle MD, Paul NS. 2015. Biochemical Marker (Protein) Based Characterization of Rice Accessions Bio-Diversity in International Rice Molecular Breeding Programme. Asian Journal of Biomedical and Pharmaceutical Sciences 5(43), 31-37.

Dudwadkar S, Parab M, Singh S. 2015. Diversity analysis among few cucurbitaceae using seed protein profile. International Journal of Plant Animal and Environmental Sciences 5(1), 146-151.

Gepts P, Bliss FA. 1988. Dissemination pathway of common bean deduced from phaseolin electrophoretic variability. Europe and Africa Economic Botany 42, 86-104.

Hossain MM, Li X, Evans IH, Rahman MA. 2014. A proteomic analysis of seed proteins expressed in a Brassica somatic hybrid and its two parental species. Plant Tissue Culture and Biotechnology 24(1), 11-26.

Iqbal SH, Ghafoor A, Ayub N. 2005. Relationship between SDS-PAGE markers and Ascochyta blight in chickpea. Pakistan Journal of Botany 37, 87-96.

Isemura T, Shiyo N, Shigeyuki M, Michihiro Y, Hiroo N, Masayoshi I, Osamu K. 2001. Genetic variation and geographical distribution of Azuki bean (Vigna angularis) landraces based on the electrophoregram of seed storage proteins. Breeding Science 51, 225-230.

Javid A, Ghafoor A, Anwar R. 2004. Seed storage protein electrophoresis in groundnut for evaluating genetic diversity. Pakistan Journal of Botany 36, 25-29.

Jiang, S, Liu S, Zhao C, Wu C. 2016. Developing protocols of Tricine-SDS-PAGE for separation of polypeptides in the mass range 1-30 kDa with minigel electrophoresis system. International Journal of Electrochemical Sience 11, 640-649.

Khan SA, Iqbal J, Khurshid H, Zia M, Shinwari ZK, Rabbani MA. 2014. Intra-specific genetic divergence in rapeseed (Brassica napus L.) genotypes estimated through SDS-PAGE of total seed proteins. International Journal of  Basic and Applied Sciences 3(2), 110-117.

Nawaz S, Chaudhry Z, Bibi A, Jan SA, Bibi K, Asma. 2015. Agro-morphological and molecular characterization of local tomato cultivars grown in Pakhal region of Pakistan using RAPD markers. Middle-East Journal of Scientific Research, 23(5), 856-860. DOI: 10.5829/idosi.mejsr.2015.23.05.9375

Rahman MM, Hirata Y. 2004. Genetic diversity in Brassica species using SDS-PAGE analysis. Journal of Biological Sciences 4, 234-238.

Rabbani MA, Qureshi AA, Afzal M, Anwar R, Komatsu S. 2001. Characterization of Mustard [Brassica juncea (L.) Czern. & Coss.] Germplasm by SDS-PAGE of Total Seed Protiens. Pakistan Journal of Botany 33(2), 173-179.

Semagn KA, Bjornstad, Ndjiondjop MN. 2006. An overview of molecular marker methods for plants. African Journal of Biotechnology 5(25), 2540-2568.

Shinwari S, Akber F, Rabbani MA, Mumtaz AS, Shinwari ZK. 2013. Evaluation of genetic diversity in different genotypes of Eruca sativa from Pakistan by SDS-PGE analysis.  Pakistan Journal of Botany 45(4), 1235-1240.

Sneath PH, Sokal RR. 1973. Numerical Taxonomy; The Principles and Practice of numerical Classification. W.F. Freeman & Co.., an Francisco, USA. 573.

Turi NA, Farhatullah, Rabbani MA, Khan NU, Akmal M, Pervaiz ZH, Aslam MU. 2010. Study of total Seed Storage Protein in Indigenous Brassica species based on Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE). African Journal of Biotechnology 9(45), 7595-7602.

Wadood SF, Hassan N, Khaliq A, Nausheen, jan T, Ghafoor A, Khan M, Nisar M. 2016. Genetic polymorphism in Lens culinaris collected from Malakand division Khayber Pakhtunkhwa, Pakistan. Journal of Biodiversity and Environmental Sciences 8(2), 53-60.

Zahoor M, Nisar M, Islam N. 2015. Genetic variations of Robinia pseudoacacia plant using SDS-PAGE. Pakistan Journal of Botany 46(6), 2335-2338.

Zada M, Shinwari ZK, Zakir N, Rabbani MA. 2013. Study of total seed storage proteins in Ethiopian mustard (Brassica carinataa. Braun) Germplasm. .  Pakistan Journal of Botany 45(2), 443-448.

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