Differentially expressed, abundant trans-spliced cDNAs from larval Brugia malayi

Mol Biochem Parasitol. 1997 Jul;87(1):85-95. doi: 10.1016/s0166-6851(97)00050-9.

Abstract

Isolation and cloning of abundant reverse transcriptase-polymerase chain reaction (RT-PCR) products from the filarial nematode Brugia malayi using the conserved nematode spliced leader sequence and poly A as amplification targets has allowed us to identify abundant, stage specific transcripts from infective and post-infective larvae. The predicted protein products of the most prominent full-length transcripts from mosquito-derived L3 parasites are: (i) Bm-ALT-1, a homologue of a Dirofilaria immitis abundant larval protein: (ii) Bm-CPI-1, a cystatin-type cysteine protease inhibitor; (iii) Bm-ALT-3, a novel predicted 6 kDa glycine/tyrosine-rich protein; and (iv) Bm-TPH-1, a homologue of a mammalian translationally-controlled tumour protein. Some transcripts were not full-length but had mis-primed at A-rich stretches of coding sequence: the most abundant of these was Bm-col-3, a which encodes a collagen homologous to Bp-COL-1 of Brugia pahangi. Similar analysis of abundant spliced leader (SL)/oligo-dT products from fourth-stage larvae 9 days post-infection yielded two dominant transcripts: (i) Bm-cdd-1, which encodes a protein with homology to cytidine deaminase, differing at only one amino acid position from its homologue described in Brugia pahangi; and (ii) the same truncated form of Bm-col-3 found in L3 preparations. Expression of the major transcripts was assessed by PCR amplification of cDNA libraries derived from each stage of the life cycle. alt1, alt-3 and cpi-1 were all found to be specific to the L3 stage, while cdd-1 was found only in the L4 cDNA library. Expression of these larval-specific transcripts was not detected in either microfilarial or adult libraries.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aedes / parasitology
  • Amino Acid Sequence
  • Animals
  • Brugia malayi / genetics*
  • Consensus Sequence
  • DNA, Complementary / genetics*
  • DNA, Helminth / genetics*
  • Gene Expression Regulation*
  • Gene Library
  • Gerbillinae
  • Larva
  • Molecular Sequence Data
  • Polymerase Chain Reaction
  • RNA Splicing
  • RNA, Helminth / genetics*
  • RNA, Helminth / metabolism
  • RNA, Messenger / genetics*
  • RNA, Messenger / metabolism
  • Sequence Alignment
  • Sequence Homology, Amino Acid

Substances

  • DNA, Complementary
  • DNA, Helminth
  • RNA, Helminth
  • RNA, Messenger

Associated data

  • GENBANK/D84432
  • GENBANK/N44411
  • GENBANK/R47601
  • GENBANK/R95314
  • GENBANK/U29459
  • GENBANK/U29576
  • GENBANK/U29577
  • GENBANK/U29578
  • GENBANK/U57547
  • GENBANK/U61949
  • GENBANK/U64861
  • GENBANK/U80971
  • GENBANK/U80972
  • GENBANK/U80973
  • GENBANK/U80974
  • GENBANK/U80975
  • GENBANK/U80976
  • GENBANK/U80977
  • GENBANK/U80978
  • GENBANK/U80979
  • GENBANK/U80980
  • GENBANK/U81008
  • GENBANK/U84723
  • GENBANK/U84736
  • GENBANK/X16064
  • GENBANK/X95663
  • GENBANK/Z28056
  • GENBANK/Z37160
  • GENBANK/Z79754