Molecular cloning and immunolocalization of the 17 kDa myoglobin of Clonorchis sinensis.
We purified the 17 kDa protein abundant in Clonorchis sinensis crude extracts. The N-terminal amino acid sequence of this protein was determined and an oligonucleotide probe synthesized. Using this probe, the cDNA encoding the protein was cloned and sequenced from the C. sinensis cDNA library. It was found to consist of a total of 150 amino acids and to have 41% conserved homology with the myoglobin of the trematodes Paramphistomum epiclitum and Isoparorchis hypselobagri. The gene product over-expressed in the bacterial system was purified and identified as the same molecule in the adult worms. BALB/c mouse sera raised against the adult 17 kDa protein revealed that this myoglobin was distributed throughout the parenchymal tissues except for the eggs and reproductive organs and that the protein may be involved in the survival of C. sinensis in the oxygen-depleted environment of the host.
Department of Parasitology, College of Medicine, Cheju National University, Jeju 690-756, Korea.
Young-Bae
YB
Chung
Hyun-Jong
HJ
Yang
Sung-Jong
SJ
Hong
Shin-Yong
SY
Kang
Mejeong
M
Lee
Tae Yun
TY
Kim
Min-Ho
MH
Choi
Jong-Yil
JY
Chai
Sung-Tae
ST
Hong
D000595:Amino Acid Sequence; D000818:Animals; D000947:Antigens, Helminth / Q000276:immunology; D003001:Cloning, Molecular; D003004:Clonorchis sinensis / Q000737:chemistry* / Q000235:genetics; D018076:DNA, Complementary; D015723:Gene Library; D015801:Helminth Proteins / Q000032:analysis* / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D007150:Immunohistochemistry; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D009211:Myoglobin / Q000032:analysis* / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D010100:Oxygen / Q000378:metabolism; D011994:Recombinant Proteins / Q000032:analysis / Q000737:chemistry / Q000276:immunology; D017386:Sequence Homology, Amino Acid
D000947:Antigens, Helminth; D018076:DNA, Complementary; D015801:Helminth Proteins; D009211:Myoglobin; D011994:Recombinant Proteins; D010100:Oxygen