Analysis of membrane protein genes in a Brazilian isolate of Anaplasma marginale
Analysis of membrane protein genes in a Brazilian isolate of Anaplasma marginale
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DOI：
10.1590/s0074-02762010000700001
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发表时间：
2010-11-01
期刊：
影响因子：
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通讯作者：
Fonseca, Adivaldo H
中科院分区：
文献类型：
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作者：
SG Junior, Daniel;Araújo, Flábio R;Fonseca, Adivaldo H
对边缘无浆体全基因组的测序使人们能够识别几个编码膜蛋白的基因，从而增加了识别候选免疫原的机会。关于编码边际疟原虫分离株中膜蛋白的基因的遗传变异性，人们知之甚少。本研究的目的是确定OMP1、OMP4、OMP5、OMP7、OMP8、OMP10、OMP14、OMP15、SODb、OPAG1、OPAG3、VirB3、VirB9-1、PepA、EF-Tu和AM854蛋白的预测氨基酸序列与其他分离株相比的保守程度。因此，我们用引物扩增了这些基因：omp1、omp4、omp5、omp7、omp8、omp10、omp14、omp15、sodb、opag1、opag3、virb3、VirB9-1、Pepa、ef-tu和am854。聚合酶链式反应扩增后，用Sanger方法克隆和测序，用CLUST-ALW和MEGA-4程序对预测的氨基酸序列进行多比对，比较巴西株、圣马里斯株、佛罗里达州株和边缘支原体的预测序列。除外膜蛋白(OMP)7外，所有蛋白与其他边际念珠菌的同源性为92-100%。然而，只有OMP1、OMP5、EF-Tu、VirB3、SODb和VirB9-1被选为能够促进分离株之间交叉保护的潜在免疫原，因为与A.(中心)边缘也有很高的同源性(超过72%)。
The sequencing of the complete genome of Anaplasma marginale has enabled the identification of several genes that encode membrane proteins, thereby increasing the chances of identifying candidate immunogens. Little is known regarding the genetic variability of genes that encode membrane proteins in A. marginale isolates. The aim of the present study was to determine the degree of conservation of the predicted amino acid sequences of OMP1, OMP4, OMP5, OMP7, OMP8, OMP10, OMP14, OMP15, SODb, OPAG1, OPAG3, VirB3, VirB9-1, PepA, EF-Tu and AM854 proteins in a Brazilian isolate of A. marginale compared to other isolates. Hence, primers were used to amplify these genes: omp1, omp4, omp5, omp7, omp8, omp10, omp14, omp15, sodb, opag1, opag3, virb3, VirB9-1, pepA, ef-tu and am854. After polimerase chain reaction amplification, the products were cloned and sequenced using the Sanger method and the predicted amino acid sequence were multi-aligned using the CLUST-ALW and MEGA 4 programs, comparing the predicted sequences between the Brazilian, Saint Maries, Florida and A. marginale centrale isolates. With the exception of outer membrane protein (OMP) 7, all proteins exhibited 92-100% homology to the other A. marginale isolates. However, only OMP1, OMP5, EF-Tu, VirB3, SODb and VirB9-1 were selected as potential immunogens capable of promoting cross-protection between isolates due to the high degree of homology (over 72%) also found with A. (centrale) marginale.