Analysis of partial cDNA sequence from Theileria sergenti

Park, Jin-ho; Chae, Joon-seok; Kim, Dae-hyuk; Jang, Yong-suk; Kwon, Oh-deog; Lee, Joo-mook

Korean Journal of Veterinary Research 1999;39(4):797-805.

Jin-ho Park¹, Joon-seok Chae², Dae-hyuk Kim³, Yong-suk Jang³, Oh-deog Kwon¹, Joo-mook Lee¹

¹College of Veterinary Medicine, Chonbuk National University
²School of Veterinary Medicine
³Faculty of Biological Sciences, Chonbuk National University

Abstract

T sergenti cDNA library were constructed to get a more broad information about the structural, functional or antigenic properties of the proteins, and analyzes for their partial cDNA sequences and expression sequences tags(ESTg). The mRNA were purified from T sergenti isolates to identify the information of antigen gene, then first and second strand cDNA was synthesized. EcoR I adaptor ligation and Xho I enzyme restriction were used to the synthesized cDNA, and ligated into a Uni-ZAP XR vector. T sergenti cDNA library was constructed with packaging and amplification in vitro. Antibody screening was performed with constructed T sergenti cDNA library using antisera against T sergenti. Among those clones, eight phagemids were rescued from the recombinant in vivo excision with f1 helper phage. Using the analysis of endonuclease restriction and PCR, the recombinant cDNA were proved having a 0.5-3.0kb of inserts. The eight of partial cDNA clones' sequences were obtained and examined for their homology using BLASTN and BLASTX. The eight of sequenced clones were classified into three groups according to the basis of database searches. A total 3,045bp of partial cDNA sequence were determined from six clones. The putatively identified clones contain a cytochrome c gene, a heat shock protein gene, a cyclophilin gene, and a ribosomal protein gene. The unidentified clones have a homology to ATP-binding protein(mtrA) gene of S argillaceus, DNA-binding protein(DBP) gene of Pseudorabies virus 85kDa merozoite protein gene of B bovis, mRNA spm1 protein of T annulata and glycine-rich RNA-binding protein mRNA of O sativa etc.

Key Words: Theileria sergenti, cDNA library, expression sequences tags(ESTg)