Construction and initial analysis of a representative lambda ZAPII expression library of the intracellular rickettsia Cowdria ruminantium: cloning of map1 and three other Cowdria genes

被引:18
作者
Brayton, KA [1 ]
Fehrsen, J [1 ]
deVilliers, EP [1 ]
vanKleef, M [1 ]
Allsopp, BA [1 ]
机构
[1] ONDERSTEPOORT VET INST,ZA-0110 ONDERSTEPOORT,SOUTH AFRICA
关键词
Cowdria ruminantium; immunoaffinity; expression library; map1; gene; genetics;
D O I
10.1016/S0304-4017(97)00020-4
中图分类号
R38 [医学寄生虫学]; Q [生物科学];
学科分类号
07 ; 0710 ; 09 ; 100103 ;
摘要
The causative agent of heartwater, the rickettsia Cowdria ruminantium, is very poorly understood at the molecular level owing to a profound lack of suitable tools. We have developed an immunoaffinity chromatographic method to purify C. ruminantium from host cell components and the purified rickettsial cells have been used to prepare substantially pure Cowdria DNA. This DNA has been used to construct what we believe to be the first fully representative C. ruminantium expression library. A clone containing the complete Cowdria map1 gene has been isolated and sequenced. This gene has been expressed in E. coli cells from the native Cowdria promoter, suggesting that the mechanisms for gene transcription and translation are similar between these two organisms. Parts of three other Cowdria genes have also been isolated and sequenced. (C) 1997 Elsevier Science B.V.
引用
收藏
页码:185 / 199
页数:15
相关论文
共 50 条
[1]   MOLECULAR-BASIS FOR SURFACE-ANTIGEN SIZE POLYMORPHISMS AND CONSERVATION OF A NEUTRALIZATION-SENSITIVE EPITOPE IN ANAPLASMA-MARGINALE [J].
ALLRED, DR ;
MCGUIRE, TC ;
PALMER, GH ;
LEIB, SR ;
HARKINS, TM ;
MCELWAIN, TF ;
BARBET, AF .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1990, 87 (08) :3220-3224
[2]  
ALLSOPP BA, 1996, ANN NY ACAD SCI, V971, P17
[3]  
ALTSCHUL SF, 1990, J MOL BIOL, V215, P403, DOI 10.1006/jmbi.1990.9999
[4]  
AMBROSIO RE, 1987, ONDERSTEPOORT J VET, V54, P255
[5]  
[Anonymous], 1994, PROGRAM MANUAL WISCO
[6]  
BANKIER AT, 1987, METHOD ENZYMOL, V155, P51
[7]   THE MSP1-BETA MULTIGENE FAMILY OF ANAPLASMA-MARGINALE - NUCLEOTIDE-SEQUENCE ANALYSIS OF AN EXPRESSED COPY [J].
BARBET, AF ;
ALLRED, DR .
INFECTION AND IMMUNITY, 1991, 59 (03) :971-976
[8]  
BARRE N, 1987, ONDERSTEPOORT J VET, V54, P405
[9]  
BENSON S A, 1984, Biotechniques, V2, P126
[10]  
BEZUIDENHOUT JD, 1985, ONDERSTEPOORT J VET, V52, P113