Construction of Larval cDNA Expression Library and Immunological Identification of Positive Clones in Tick Haemaphysalis qinghaiensis

文献类型： 外文期刊

第一作者： Zhao Hai-ping

作者： Zhao Hai-ping;Yin Hong;Gao Jin-liang;Li You-quan;Guan Gui-quan;Liu Zhi-jie;Liu Jun-long;Luo Jian-xun;Zhao Hai-ping;Yang Fu-he;Xing Xiu-mei

作者机构：

关键词： larval tick;Haemaphysalis qinghaiensis;cDNA expression library;immunoscreening;immunological identification

期刊名称：AGRICULTURAL SCIENCES IN CHINA （ 影响因子：0.82； 五年影响因子：0.997 ）

ISSN： 1671-2927

年卷期： 2010 年 9 卷 6 期

页码：

收录情况： SCI

摘要： A primary cDNA expression library with a titer of 5 0 x 10(5) PFU mL(-1) was constructed from mRNA extracted from larval Haemaphysalis qinghaiensis ticks in order to identify certain genes, which would then be used as candidate molecules for development of effective vaccines to control this parasite Totally 11 positive clones, which designated as HqL01-11, were obtained by immunoscreening of the library using a polyclonal antibody generated in rabbit with larval tick protein extract Results of sequence analysis from BLASTN searching revealed that 6 of them had no significant homology with the adult If. qinghaiensis ticks' known genes, 4 of them had no significant homology with all genes deposited in GenBank database HqL07, HqL08, HqL09, and HqL11 were deposited to GenBank database, and accession numbers were EF605263, EF605264, EF605265, and EF605266, respectively Subsequently, HqL07 and HqL09 were expressed in vitro and the molecular weights of the corresponding expressed products were 60 and 70 kDa, respectively. Western blot analyses showed that HqL07 and HqL09 had immunogenicity This study laid the foundation for future production of genetically engineered vaccines for the immunological control of H. quighaiensis

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