RAPD-PCR based genomic characterization of two populations of Culex quinquefasciatus (Diptera : Culicidae)

Abstract

Abstract:The present paper deals with the RAPD-PCR based genomic characterization of Culex quinquefasciatus Say which is a major vector of filariasis in several parts of the Indian subcontinent. One population of the test organism used in the study was procured from Goa (pop.A) while the other (pop.B) was collected from a village Nadasahib (20 kms from Chandigarh). The RAPD-PCR amplification of whole body homogenate of freshly hatched individual specimens was carried out by using three random primers: primer I-5'-GTCCCGACGA -3'; primer II-5' -TGATCCCTGG -3' and primer III-5'-GTGACGTAGG -3'. Primer I produced 5 distinct bands from the DNA of pop. A, whose base composition ranged from 200-1000 bp. Likewise, 7 bands ranging from 130-750 bp and 4 bands ranging from 270-950 bp were generated with primers II and III respectively. In case of pop.B, a total of 8 bands ranging from 200-1000 bp were generated with primer I. Similarly, a total of 6 bands ranging from 250-900 bp with primer II and 4 bands ranging from 180-950 bp with primer III were produced. Based on the band sharing coefficient and the application of Nearest Neighbour Joining (NJ) analysis it was found that primer I was more suitable for detecting genomic differences at the species and generic levels while primer II was ideal for detecting variations in the number of bp in RAPD generated bands among different populations of Cx. quinquefasciatus.

Genome fingerprinting of two populations of Aedes (Stegomyia) albopictus Skuse (Asian Tiger) using random primers (Diptera: Culicidae)

“…It is also logical to state that differences in their DNA profiles are attributed to the spacial segregation which tends to influence the genotype qualities of populations. The changes or the variations at the genome level have produced differences in the RAPD profiles which also revealed the genome as a source of consta ntly changing sequences among the populations and vector (Kengne et al, 2001;Kaura et al, 2009). The classic works on the distribution, bionomics, epidemiology and genetics of this taxon provide enough impetus to analyse the genome organization of as many populations of the species as possible to understa nd the patterns of sequence va riations (Narang et al, 1993;Mitchell, 1995;Ayres et al, 2002).…”

“…Hamirpur Himachal Pradesh (Latitude: 76º 18' -76º 44' E; Longitude: 31º 25' to 31º 52' N) while the other (pop.B) was procured from Panjab University campus, Chandigarh (Latitude: 30°43'N; Longitude: 76°47'E). The genomic DNA was extracted from the legs of individual specimen by following a standard phenol-chloroform extraction method with minor modifications (Ausubel et al, 1999;Chaudhry, 1999;Kaura et al, 2009). The purified and quantified samples of DNA were amplified by using three random primers viz: Primer I-5'-TTTGCCCGGA-3', Primer II-5'-GTCCCGACGA-3' and Primer III-5'-CAGGCCCTTC-3'.…”