Characterization and immunolocalization of a Cryptosporidium protein containing repeated amino acid motifs
Characterization and immunolocalization of a Cryptosporidium protein containing repeated amino acid motifs
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含有重复氨基酸基序的隐孢子虫蛋白的表征和免疫定位
DOI：
10.1128/iai.61.6.2347-2356.1993
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发表时间：
1993
影响因子：
3.1
通讯作者：
A. Crisanti
中科院分区：
文献类型：
--
作者：
L. Ranucci;H. Müller;G. La Rosa;I. Reckmann;M. A. Morales;F. Spano;E. Pozio;A. Crisanti
卵囊壁是使隐孢子虫在环境中存活并保持感染性的成分之一。为了鉴定在卵囊阶段特异性表达的隐孢子虫蛋白，我们用卵囊抗血清和特异性基因探针筛选了微小隐孢子虫的λ gt 11基因组文库。我们从不同的文库中分离出两个重叠克隆，它们含有编码1，252个氨基酸多肽的开放阅读框。推导的氨基酸序列的分析显示异常高的含量的半胱氨酸，脯氨酸和组氨酸。该序列的特征还在于两个不同的氨基酸基序，每个重复多次。编码氨基酸重复序列的DNA序列显示出高频率的同义突变，结果表明重复的基序可能在功能上和/或结构上对寄生虫很重要。针对包含前786个氨基酸的重组多肽开发的抗血清和单克隆抗体显示，C. parvum的表观分子量为190，000。此外，免疫荧光共聚焦显微镜分析表明，蛋白质定位在卵囊壁作为一个统一的染色和卵囊本身作为明亮的颗粒与残留的身体密切相关。
The oocyst wall is one of the components that permits cryptosporidia both to survive in the environment and to retain infectivity. With the aim of identifying Cryptosporidium proteins specifically expressed at the oocyst stage, we screened lambda gt11 genomic libraries of Cryptosporidium parvum with both an oocyst antiserum and a specific genetic probe. We isolated, from distinct libraries, two overlapping clones containing an open reading frame encoding a 1,252-amino-acid polypeptide. The analysis of the deduced amino acid sequence revealed unusually high contents of cysteine, proline, and histidine. The sequence was also characterized by two distinct amino acid motifs, each repeated several times. The DNA sequences coding for the amino acid repeats showed a high frequency of synonymous mutations, a result suggesting that the repeated motifs may be functionally and/or structurally important to the parasite. Antisera and monoclonal antibodies developed against a recombinant polypeptide encompassing the first 786 amino acids revealed that the corresponding protein in C. parvum had an apparent molecular weight of 190,000. Moreover, confocal microscopy analysis with immunofluorescence indicated that the protein was localized on the oocyst wall as a uniform stain and within the oocyst itself as bright granules in close association with the residual body.
小隐孢子虫肌动蛋白基因的分离、序列和分子核型分析。
影响因子：
1.5
作者：
Kim,K;Goozé,L;Petersen,C;Gut,J;Nelson,RG
通讯作者：
Nelson,RG