Hammoudeh Nour, Kweider Mahmoud, Abbady Abdul-Qader, Soukkarieh Chadi
Dept. of Biotechnology, Faculty of Agriculture, Damascus University, Damascus, Syria.
Dept. of Animal Biology, Faculty of Sciences, Damascus University, Damascus, Syria.
Iran J Parasitol. 2014 Oct-Dec;9(4):574-83.
Leishmania Homologue of receptors for Activated C Kinase (LACK) antigen is a 36-kDa protein, which provokes a very early immune response against Leishmania infection. There are several reports on the expression of LACK through different life-cycle stages of genus Leishmania, but only a few of them have focused on L.tropica.
The present study provides details of the cloning, DNA sequencing and gene expression of LACK in this parasite species. First, several local isolates of Leishmania parasites were typed in our laboratory using PCR technique to verify of Leishmania parasite species. After that, LACK gene was amplified and cloned into a vector for sequencing. Finally, the expression of this molecule in logarithmic and stationary growth phase promastigotes, as well as in amastigotes, was evaluated by Reverse Transcription-PCR (RT-PCR) technique.
The typing result confirmed that all our local isolates belong to L.tropica. LACK gene sequence was determined and high similarity was observed with the sequences of other Leishmania species. Furthermore, the expression of LACK gene in both promastigotes and amastigotes forms was confirmed.
Overall, the data set the stage for future studies of the properties and immune role of LACK gene products.
活化C激酶受体的利什曼原虫同源物（LACK）抗原是一种36 kDa的蛋白质，可引发针对利什曼原虫感染的非常早期的免疫反应。关于LACK在利什曼原虫属不同生命周期阶段的表达已有多篇报道，但其中只有少数聚焦于热带利什曼原虫。
本研究详细介绍了该寄生虫物种中LACK的克隆、DNA测序和基因表达情况。首先，在我们实验室使用PCR技术对几种利什曼原虫的本地分离株进行分型，以验证利什曼原虫的物种。之后，扩增LACK基因并克隆到载体中进行测序。最后，通过逆转录PCR（RT-PCR）技术评估该分子在对数生长期和稳定期前鞭毛体以及无鞭毛体中的表达。
分型结果证实我们所有的本地分离株均属于热带利什曼原虫。确定了LACK基因序列，并观察到与其他利什曼原虫物种的序列具有高度相似性。此外，证实了LACK基因在前鞭毛体和无鞭毛体形式中的表达。
总体而言，这些数据为未来研究LACK基因产物的特性和免疫作用奠定了基础。
