Cloning and characterization of a repetitive DNA sequence specific for Wuchereria bancrofti.
A genomic library constructed in a bacteriophage lambda replacement vector (EMBL3) with Wuchereria bancrofti DNA partially digested with Sau 3A I was screened with 32P-labeled W. bancrofti total DNA, and a strongly reactive recombinant, EMBL3Wb34, was isolated. This clone contained an approximately 16-kb insert that showed some cross-hybridization with Brugia malayi and B. pahangi DNA. However, a 969-bp subclone from EMBL3Wb34, designated pWb12, hybridized only with W. bancrofti DNA and was able to detect as little as 300 pg. Furthermore, pWb12 could detect DNA from a single infective larva or one microfilaria. It has a moderate copy number (450-700) and appears to be interspersed within the parasite genome. The nucleotide sequence contains 66% A+T and 34% G+C and shows no notable internal repeats.
Siridewa
K
K
Department of Biochemistry and Molecular Biology, Faculty of Medicine, University of Colombo, Sri Lanka.
Karunanayake
E H
EH
Chandrasekharan
N V
NV
Abeyewickreme
W
W
Franzen
L
L
Aslund
L
L
Pettersson
U
U
D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D003465:Culex / Q000235:genetics; D018509:DNA, Helminth / Q000737:chemistry*; D015723:Gene Library; D005822:Genetic Vectors; D006801:Humans; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D012091:Repetitive Sequences, Nucleic Acid; D013045:Species Specificity; D013188:Sri Lanka; D014958:Wuchereria bancrofti / Q000235:genetics*
D018509:DNA, Helminth