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Taruna Kaura Ravinder Kumar Sudarshan Chaudhry

Abstract

The present paper deals with RAPD-PCR based genomic characterization of two allopatric populations of Aedes (Stegomyia) albopictus Skuse which is a major vector of dengue fever and a source of many other pathogenic infections in man. For the present purpose one population was collected from village Dhunater, Distt. Hamirpur in Himachal Pradesh (pop.A) while the other was procured from Panjab University campus (pop.B). The genomic DNA from the legs of individual specimens was amplified by using three random primers viz: Primer I-5’TTTGCCCGGA-3’, Primer II- 5’-GTCCCGACGA-3’ and Primer III- 5’-CAGGCCCTTC-3’. The amplification of the DNA of pop.A with primer I, produced a total of 7 bands ranging from 230-880 bp while 3 bands ranging from 450-820 bp were produced from pop. B. Similarly, with primer II a total of 8 bands were produced from pop. A, which ranged from 220-800 bp while 9 were produced from the DNA of the individuals of pop. B, which ranged from 200-900 bp. With primer III, a total of 15 bands were produced from pop. A, with a base pair composition varying from 210-1031 bp while 5 were produced from pop. B with a range of 210-370 bp. From the hierarichial cluster sharing analysis of bands, primers I and II were found to ideal for the differentiation of the individuals at the population level studies of this species of considerable epidemiological significance.

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Keywords

Ae, Albopictus, Allopatric populations, Genome fingerprinting, RAPD-PCR

References
Ausubel, F.M., Breut, R., Kingston, R.E., Mooer, D.D., Seidman, J.G., Smith, J.A., and Struhl, K. (1999). Short protocols in molecular biology, London: John-Wiley and Sons Inc.
Ayres, C.F.J., Romao, T.P.A, Melo-Santos, M.A.V. and Furtado, A.F. (2002). Genetic diversity in brazilian populations of Aedes albopictus. Mem Inst Ostwaldo Cruz Rio de Janeiro. 97(6): 871-875.
Ballinger- Crabtree, M.E., Black, W.C. and Miller, B.R. (1992). Use of genetic polymorphisms detected by the Random Amplified Polymorphic DNA Polymerase Chain Reaction (RAPD-PCR) for differentiation and identification of Aedes aegypti subspecies and populations. American Journal of Tropical Medicine and Hygiene. 47(6): 893-901.
Black, W.C. IV and DuTeau, N.M. (1996). RAPD-PCR and SSCP analysis for Insect population genetic studies. In: The molecular biology of insect disease vectors: a methods manual (ed J. Crampton). New York: Chapman and Hall.
Carlson, D.A. and Service, M.W. (1980). Identification of mosquitoes of Anopheles gambiae species complex A and B by analysis of cuticular components. Science. 207(35): 1089-1091.
Chaudhry, S. (1999). A review of polytene chromosome dynamics in genus Anopheles (Diptera: Culicidae) Some aspects on the insight of insect biology (eds R.C. Sobti and J.S. Yadav), pp. 205-236. Tausco Book Distributors, New Delhi.
Chaudhry, S., Sharma, M., Gupta, S. and Chillar, J.S. (2006). Multiple technique based species discrimination in the taxon Anopheles stephensi (Culicidae: Diptera). In: Proceedings of the International Symposium on Vector Biology (eds V.P. Sharma, J.S. Kirti), pp. 105-123. Panjabi University, Patiala, India.
Conn, J, Puertas, Y.R. and Seawright, J.A. (1993). A new cytotype of Anopheles nuneztovari from Western Venezuela and Colombia. Journal of American Mosquito Control Association. 9(3): 294-301.
Craven, R.B., Eliason, D.A., Francy, D.B., Reiter, P., Campos, E.G., Jakob, W. L., Smith, G.C., Bozzi, C.J., Moore, C.G., Maupin, G.O. et al. (1988). Importation of Aedes albopictus and other exotic mosquito species into the United States in used tyres from Asia. Journal of American Mosquito Control Association. 4: 138-142.
de Sousa, G.B., Blanco, A. and Gardenal, C.N. (2001). Genetic relationships among Aedes aegypti (Diptera:Culicidae) populations from Argentina using Random Amplified Polymorphic DNA Polymerase Chain Reaction markers. Journal of Medical Entomolgy. 38(3): 371-375.
dos Santos, V.M., Macoris, M.L.and Andrighetti, M.T.(2003). Analysis of genetic relatedness between populations of Aedes aegypti from different geographic regions of São Paulo state, Brazil. Revista do Instituto de Medicina Tropical de São Paulo. 45(2): 99-101
Favia, G., Dimopoulos, G., Torre, A.D., Toure, Y.T., Coluzzi, M. and Louis, C. (1994). Polymorphisms detected by random PCR distinguish between different chromosomal forms of Anopheles gambiae. Proceeding National Academy of Science USA. 91:10315-10319.
Kambhampati, S., Black IV, W.C. and Rai, K.S. (1992). Random amplified polymorphic DNA of mosquito species and populations (Diptera: Culicidae): techniques, statistical analysis and applications. Journal of Medical Entomology, 29: 939-945.
Kaura, T., Kaushal, P., Bansal, M., Chaudhry, S. and Chaudhry, A. (2009). RAPD-PCR based genomic characterization of Culex quinquefasciatus (Diptera: Culicidae). Journal of Applied Natural Science. 1(2): 269-274.
Kengne, P., Trung, H.D., Baimai, V., Coosemans, M. and Manguin, S. (2001). A multiplex PCR-based method derived from random amplified polymorphic DNA (RAPD) markers for the identification of species of the Anopheles minimus group in South-east Asia. Insect Molecular Biology. 10: 427-435.
Knudson, D.L., Zheng, L. and Gordon, S.W. (1996). Genome Organization of vectors In the Biology of Disease Vectors (eds B. J. Beaty and W. C. Marquardt), pp. 175-214. University Press of Colorado Niwot, USA.
Kumar, A. and Rai K.S. (1993). Molecular organization and evolution of mosquito genomes. Comparative Biochemistry and Physiology - Part B: Biochemistry & Molecular Biology. 106: 495-504.
Manguin, S., Kengne, P. and Sonnier, L. (2002). SCAR markers and multiplex PCR-based identification of isomorphic species in the Anopheles dirus complex in South-east Asia. Medical Veternary Entomology. 16(1): 46-54.
Mathiopoulos, K.D., Powell, J.D. and Mc Cutchan, T.F. (1995). An anchord restriction-mapping approach applied to the genetic analysis of the Anopheles gambiae malaria vector complex 1. Molecular Biology Evolution. 12: 103-112.
Mitchell, C.M. (1995). Geographic spread of Aedes albopictus and potential for involvement in arbovirus cycles in the Mediterranean basin. Journal of Vector Ecology. 20: 44-58.
Munstermann, L.E. (1995). Mosquito systematics: Current status, new trends, associated complications. Journal of Vector Ecology. 20: 129-138.
Mutebi, J.P., Black IV, W.C., Bosio, C.F., Sweeney, W.P. Jr. and Craig, G.B. Jr. (1997). Linkage map for the asian tiger mosquito (Aedes (Stegomyia) albopictus) based on SSCP analysis of RAPD markers. Journal of Heredity. 88(6): 489-494.
Narang S.K., Seawright J.A., Mitchell S.E., Kaiser, P.E. and Carlson, D.A. (1993). Multiple-technique identification of sibling species of the Anopheles quadrimaculatus complex. Journal of American Mosquito Control Association. 9(4): 463-464.
Rai, K.S. (1991). Aedes albopictus in the Americas. Annual Review of Entomology. 36: 459-484.
Rao P.N. and Rai S.K. (1988). Inter- and Intra-specific variation in nuclear DNA content in Aedes mosquitoes. Heredity. 59: 253-258.
Rudnick A. and Chan Y.C. (1965) Dengue type 2 virus in naturally infected Aedes albopictus mosquitoes in Singapore. Science. 149: 638-639.
Sambrook J., Fritsch E.F. and Maniatis T. (1989) Molecular cloning: a laboratory manual, 2nd ed. New York: Laboratory Press, Cold Spring Harbor.
Shouche Y.S. and Patole M.S. (2000) Sequence analysis of mitochondrial 16S ribosomal RNA gene fragment from seven mosquito species. Journal of Bioscience. 25(4): 361-366.
Sucharit S. and Komalamisra N. (1997) Differentiation of Anopheles minimus species complex by RAPD-PCR technique. Journal of Medical Assocciation Thailand. 80(9): 598-602.
Welsh J. and McClelland M. (1990) Genome fingerprinting using arbitrarily primed PCR and a matrix of pairwise combinations of primers. Nucleic Acids Research, 19(19): 5275-5279.
Section
Research Articles

How to Cite

Genome fingerprinting of two populations of Aedes (Stegomyia) albopictus Skuse (Asian Tiger) using random primers (Diptera: Culicidae). (2013). Journal of Applied and Natural Science, 5(1), 95-102. https://doi.org/10.31018/jans.v5i1.288