Construction and analysis of cDNA library of Necator americanus third stage larvae.
OBJECTIVE
To obtain the genetic information on Necator americanus and to search for the purpose genes.


METHODS
mRNA was isolated from the third stage larvae of Necator americanus maintained in hamsters. Double strand cDNA was synthesized and ligated to lambda ZAPII vector to construct the cDNA library. Expressed sequence tages (ESTs) were obtained by single pass sequencing of randomly isolated cDNA clones from the established library.


RESULTS
A cDNA library of N. americanus was successfully constructed with high recombinant efficiency. The titer of unamplified library was 1 x 10(7). The insert size was about 750-3,000 bp. Of 11 ESTs obtained from the library, 7 have a significant homology with certain functional genes.


CONCLUSIONS
A high quality and high representative cDNA library of N. americanus was constructed at the first time and some functional genes were identified from the library by ESTs.
Zhan
B
B
Institute of Parasitic Diseases, Chinese Academy of Preventive Medicine, WHO Collaborating Center for Malaria, Schistosomiasis and Filariasis, Shanghai, 200025.
Hawdon
J
J
Shan
Q
Q
Ren
H
H
Qiang
H
H
Xiao
S H
SH
Li
T H
TH
Feng
Z
Z
Hotez
P
P
D000818:Animals; D006224:Cricetinae / Q000469:parasitology; D018076:DNA, Complementary / Q000235:genetics*; D020224:Expressed Sequence Tags; D015723:Gene Library; D007814:Larva / Q000235:genetics; D017221:Necator americanus / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics / Q000302:isolation & purification
D018076:DNA, Complementary; D012333:RNA, Messenger