TY - JOUR
T1 - Genes expressed in Brugia malayi infective third stage larvae
AU - Blaxter, Mark L.
AU - Raghavan, Nithyakalyani
AU - Ghosh, Inca
AU - Guiliano, David
AU - Lu, Wenhong
AU - Williams, Steven A.
AU - Slatko, Barton
AU - Scott, Alan L.
N1 - Funding Information:
This work was supported by researchg rants from the UNDP/World Bank/World Health OrganisationS pecial Programmef or Researcha nd Training in Tropical Diseases,U S Public Health Service (AI-29411 ) , the Edna McConnell Clark Foundation (EMCF 01093)a nd the MacArthur Foundation Consortium on the Biology of Parasitic Disease,t he Johns Hopkins Program. Para-sitesa nd infectedg erbilsw ere suppliedu nder the auspices of an NIAID supply contract (AI # 02642) US/Japan CooperativeM edical Science Program. M.L. Blaxter is a Wellcome Research Career Development Fellow. We would like to thank W.F. Gregory for communicating unpublished results, R.M. Maizels for support through this project and J.B. Parkes for help in data analysis.
PY - 1996/4
Y1 - 1996/4
N2 - We have used a tag sequencing approach to survey genes expressed in the third stage infective larvae of the human filarial nematode parasite Brugia malayi. RNA was isolated from late vector-stage L3 larvae after days 9 or 10 of infection in mosquitos, and converted to cDNA by reverse transcriptase. Double-stranded cDNA was produced by either conventional methods (non-SL cDNA library) or by PCR using the nematode spliced leader (SL1) and oligo(dT) primers (SL cDNA library). Two clone libraries (one from SL and one from non-SL cDNAs) were constructed in lambda ZapII. A set of these full-length clones was selected and 596 inserts were sequenced from the 5' end. We have identified 364 malayi genes (the majority of which are new) that encode housekeeping proteins, structural proteins, proteins of immediate immunological or drug-discovery interest as well as a large class of novel sequences which may prove to have significant involvement in host invasion. Extensive, genome-wide approaches to the analysis of larval gene expression are now possible for B. malayi. We present several examples of this approach.
AB - We have used a tag sequencing approach to survey genes expressed in the third stage infective larvae of the human filarial nematode parasite Brugia malayi. RNA was isolated from late vector-stage L3 larvae after days 9 or 10 of infection in mosquitos, and converted to cDNA by reverse transcriptase. Double-stranded cDNA was produced by either conventional methods (non-SL cDNA library) or by PCR using the nematode spliced leader (SL1) and oligo(dT) primers (SL cDNA library). Two clone libraries (one from SL and one from non-SL cDNAs) were constructed in lambda ZapII. A set of these full-length clones was selected and 596 inserts were sequenced from the 5' end. We have identified 364 malayi genes (the majority of which are new) that encode housekeeping proteins, structural proteins, proteins of immediate immunological or drug-discovery interest as well as a large class of novel sequences which may prove to have significant involvement in host invasion. Extensive, genome-wide approaches to the analysis of larval gene expression are now possible for B. malayi. We present several examples of this approach.
KW - Brugia malayi, third stage larva
KW - Expressed sequence tag
KW - Nematode
KW - Polymerase chain reaction
KW - Spliced leader
KW - Trans-splicing
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U2 - 10.1016/0166-6851(96)02571-6
DO - 10.1016/0166-6851(96)02571-6
M3 - Article
C2 - 8784774
AN - SCOPUS:0029989092
SN - 0166-6851
VL - 77
SP - 77
EP - 93
JO - Molecular and Biochemical Parasitology
JF - Molecular and Biochemical Parasitology
IS - 1
ER -