[Cloning and fusion expression of adenosine deaminase of Schistosoma japonicum].
OBJECTIVE
To clone and express a new protein adenosine deaminase of Schistosoma japonicum (SjADA).


METHODS
Specific primers were designed according to the EST sequence and used for amplification of the encoding sequence from the cDNA clone containing SjADA. The gene was sub-cloned into pET32 plasmid and expressed in E.coli BL31 (DE) pLys strain and the recombinant proteins were purified by chelating sepharose FF. The structure and phylifunctions of this protein were analyzed by MrBayes and GeneAtlas.


RESULTS
The full length sequence of SjADA gene was obtained and the recombinant protein was cloned, expressed and purified successfully. The bioinformatics analysis showed that the identity of SjADA and SmADA gene sequence was only 25% and they belonged to different subfamily. The structure of SjADA had 41% identity with it's PDB template 1A4M.


CONCLUSIONS
The recombinant protein of SjADA has been obtained. The purine salvage pathway of S. japonicum may be different with that of S. mansoni.
Yang
Zhong
Z
State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China Uiniversity of Science and Technology, Shanghai 200237, China.
Xu
Bin
B
Wang
Wei
W
Feng
Zheng
Z
Wei
Dong-Zhi
DZ
Hu
Wei
W
D000243:Adenosine Deaminase / Q000145:classification / Q000235:genetics* / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D020134:Catalytic Domain; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D015971:Gene Expression Regulation, Enzymologic; D015801:Helminth Proteins / Q000145:classification / Q000235:genetics* / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010802:Phylogeny; D010957:Plasmids / Q000235:genetics; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D012549:Schistosoma japonicum / Q000201:enzymology*; D017422:Sequence Analysis, DNA; D020539:Sequence Analysis, Protein
D018076:DNA, Complementary; D015801:Helminth Proteins; D011993:Recombinant Fusion Proteins; D000243:Adenosine Deaminase