[Cloning and analysis of the promoter region of Schistosoma japonicum 8k CaBP gene].
OBJECTIVE
To clone and analyze the promoter sequence of Schistosoma japonicum 8k Calcium binding protein(CaBP) (Sj8CaBP) gene.


METHODS
The genomic DNA was extracted from S. japonicum adult worms. The DNA library was constructed according to the user's manual for Clontech Universal Genome Walker(TM) Kit, and the gene-specific primer 1 and 2 (GSP1 and GSP2) of Sj8CaBP gene were designed and synthesized. GSP1 and GSP2 were coupled with the adaptor primer 1 and 2 (AP1 and AP2), then nested PCR was performed. After proper purification, the PCR product was linked to pMD-18 T vector, and the recombinant T-vector was sequenced, analyzed with the promoter prediction software and compared with the cDNA sequence of Sj8CaBP.


RESULTS
The cloned Sj8CaBP gene was about 1 079 bp in length ( GenBank accession No.AY262018), which contained a full open-reading frame with 1 intron and 2 exons. The 5'UTR has a putative promoter region including a TATA-box and a CCAAT-box. The GC-box binding region was not found in this gene, nor was that for active protein 1 (AP-1) often seen in other schistosome genes.


CONCLUSIONS
A 1 079 bp Sj8CaBP gene fragment (GenBank accession No.AY262018) was obtained from a S. japonicum genomic DNA library, containing a promoter region with TATA-box, CCAAT-box, 1 intron and 2 extrons.
Peng
Hong-juan
HJ
Department of Parasitology, First Military Medical University, Guangzhou 510515, China.
Chen
Xiao-guang
XG
Zhou
Xiao-hong
XH
Shen
Shu-Man
SM
D000818:Animals; D001483:Base Sequence; D002135:Calcium-Binding Proteins / Q000096:biosynthesis / Q000235:genetics*; D003001:Cloning, Molecular; D018509:DNA, Helminth / Q000235:genetics; D015723:Gene Library; D015801:Helminth Proteins / Q000096:biosynthesis / Q000235:genetics*; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic / Q000235:genetics*; D012549:Schistosoma japonicum / Q000235:genetics*
D002135:Calcium-Binding Proteins; D018509:DNA, Helminth; D015801:Helminth Proteins