[Cloning, sequencing of G3PD gene from Brugia malayi and prediction of B cell epitopes in its amino acid sequence].
Specific primers were designed and synthesized based on the reported glyceraldehyde-3-phosphate dehydrogenase (BmG3PD) gene of Brugia malayi (GenBank Accession No. U18137). Total RNA was extracted from Brugia malayi and its BmG3PD gene was amplified by reverse transcription-polymerase chain reaction (RT-PCR). The PCR product was purified and cloned into plasmid pGEM-T, then transformed into Escherichia coli DH5alpha. The recombinant plasmids were screened and identified by digestion with restriction enzyme and PCR amplification. The positive recombinant plasmid pGEM-T-BmG3PD was confirmed by sequencing and homology comparison. Five parameters and methods were used to predict B-cell epitopes in amino acid sequence of BmG3PD. The amplified DNA fragment (1,020 bp) had a high identity of 99% with the BmG3PD gene sequence of Brugia malayi. B-cell epitopes of BmG3PD were probably at or adjacent to 22-36, 242-255, 303-318 and 326-336 in its amino acid sequence.
Department of Parasitology, School of Basic Medical Sciences, Nantong University, Nantong 226001, China.
Dong-Fang
DF
Xie
Zheng
Z
Fang
Hai-Yan
HY
Tong
Bang-Sheng
BS
Xu
Wei-Qun
WQ
Huang
Hao
H
Fang
Qin
Q
Shen
D000818:Animals; D000947:Antigens, Helminth / Q000235:genetics* / Q000276:immunology; D017178:Brugia malayi / Q000201:enzymology / Q000235:genetics*; D003001:Cloning, Molecular; D018985:Epitopes, B-Lymphocyte / Q000235:genetics*; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000235:genetics* / Q000276:immunology; D017422:Sequence Analysis, DNA
D000947:Antigens, Helminth; D018985:Epitopes, B-Lymphocyte; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases