A study for a construction of Theileria sergenti DNA probe

Kim, Myeong-chul; Lee, Joo-mook; Kwon, Oh-deog; Chae, Joon-seok; Kim, Heung-seob

Korean Journal of Veterinary Research 1993;33(3):479-486.

Myeong-chul Kim, Joo-mook Lee, Oh-deog Kwon, Joon-seok Chae, Heung-seob Kim

Department of Veterinary Medicine, Chonbuk National University

Theileria sergenti DAN probe를 만들기 위한 기초 연구

김명철, 이주묵, 권오덕, 채준석, 김흥섭

전북대학교 수의과대학

Abstract

This study was attempted co develope a method for detection of Theileria sergenti infection on the basis of hybridization of parasite DNA with a probe. For construction of a T sergenti genomic library, T sergenti DNA was digested completely with Bam-HI and the fragments were ligated into the Bam-HI site of pUC-19 before transformation of Escherichia colistrain JM83. To detect clones containing the parasite's DNA sequences, a genomic DNA library of T sergenti constructed in pUC-19 was screened by cracking and Southern hybridization. Seven colonies were chosen from 29 colonies which were screened by transformation of Escherichia coli strain JM83. Seven transformants were comfirmed from seven colonies by cracking. The sizes of transformants were about 5Kb, 5.7Kb, 4.3Kb, 7.75Kb, 7.85Kb, 5.8Kb, 3.8Kb, respectively. DNA inserts, T sergenti DNA, and bovine DNA were hybridized with radio-labelled T sergenti DNA. Two($pT_1$, $pT_1$) of the seven inserts and T sergenti DNA reacted strongly but another 5 inserts and bovine DNA showed weak reation. All of the DNA inserts were not reaction, but T sergenti DNA were very weakly and bovine DNA were strongly reacted to hybridization with radio-labelled bovine DNA. Therefore, we obtained total 7 T sergenti DNA fragments in this study.

Key Words: Boovine, Theileria sergenti, DNA probe