MOLECULAR GENETIC VARIATIONS OF DIFFERENT POPULATIONS Of AEDES C4SPJUS(PALLAS) FROM DIFFERENT GEOGRAPHICAL HABITATS.

Abstract

Genetic variability was examined for five populations of Aedes (Ochlerotatus). • caspius (Pallas) representing different geographical isolated areas of Egypt; Fayoum, Ismailia, Qalioubiya, Red sea and Sinai. Two Molecular genetic techniques were used; firstly, Random Amplified Polymorphic DNA Polymerase Chain Reaction (RAPD-PCR). A screen of 19 random decamer oligonucleotide primers revealed three primers OPA-01, -04 and -09 which were chosen to discriminate the population frequencies of RAPD fragments. The three primers produced thirteen unique and shared bands which enable the differentiation of genetically distinct Ae. caspius populations. Secondly, variations in the nucleotide sequences of the second internal transcribed spacer (ITS2) ofnuclear ribosomal DNA (rDNA) were used based on the PCR to distinguish the five Ae. caspius populations. Two specific primers of28 oligonucleotides were used; the forward primer CP-PlA was derived from a sequence in the 5.8S RNA gene and CP-PlB primer derived from a sequence in the 28S RNA gene. Amplified DNA fragments used as a template with

labeled deoxynucleotides (dNTPs) through the direct sequence and as an insert through the cloning technique. Ma'ny clones of ITS2 of 5 Ae. caspius populations are obtained using two vectors; P Blue Script II sk (+) plasmid and T-Easy vector. Sequence analysis of ITS2 clones revealed wide genetic diversity between Ae. caspius populations.