CHARACTERIZATION OF CDNA CLONES ENCODING A MAJOR MICRONEME ANTIGEN OF SARCOCYSTIS-MURIS (APICOMPLEXA) CYST MEROZOITES

被引:24
作者
ESCHENBACHER, KH
KLEIN, H
SOMMER, I
MEYER, HE
ENTZEROTH, R
MEHLHORN, H
RUGER, W
机构
[1] RUHR UNIV BOCHUM, ARBEITSGRP MOLEK GENET, D-44780 BOCHUM 1, GERMANY
[2] RUHR UNIV BOCHUM, LEHRSTUHL SPEZIELLE ZOOL & PARASITOL, W-4630 BOCHUM, GERMANY
[3] RUHR UNIV BOCHUM, INST PHYSIOL CHEM, PROT SEQUENZIERUNGS LAB, W-4630 BOCHUM, GERMANY
[4] UNIV BONN, INST ZOOL, BONN, GERMANY
关键词
COCCIDIA; SARCOCYSTIS MURIS; MICRONEME; CDNA SEQUENCE; CYSTEINE-RICH; KALLIKREIN;
D O I
10.1016/0166-6851(93)90174-V
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Two monoclonal antibodies directed against a microneme antigen of Sarcocystis muris cyst merozoites (16/17 kDa band doublet) were used to isolate cDNA clones from a lambda ZAP expression library. Restriction analysis revealed that the inserts were highly similar, with sizes ranging between 1.8 and 2.3 kb. In addition, a full-length cDNA insert of 2.6 kb was obtained by hybridization screening. On Northern blots, a single mRNA species of 2.7 kb was detected by a cDNA-derived probe. Southern blot hybridization suggests that the gene is present as a single copy. The nucleotide sequence of the full-length clone contains a single reading frame with a coding capacity of 26.5 kDa. The hypothetical polypeptide consists of a putative N-terminal signal peptide followed by a hydrophilic domain of unknown function, and the mature protein sequence. After purifying the 16/17 kDa antigen from cyst merozoites, a partial N-terminal amino acid sequence was obtained. Thus, the identity of the cDNA sequence was confirmed. The deduced sequence of the mature protein is predominantly hydrophilic and rich in cysteine (8.7%). Database searching suggested weak homologies of the hypothetical polypeptide to plasma kallikrein, tenascin and blood coagulation factors.
引用
收藏
页码:27 / 36
页数:10
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