捻转血矛线虫脂肪酶基因表达与生物学特性分析

doi:10.11843/j.issn.0366-6964.2018.03.015

Abstract

Abstract:

To study the biological character of Lipase from Haemonchus contortus, the lipase gene of H. contortus preserved in our laboratory was cloned by PCR, and then expressed in procaryotic expression system. The expression vector was constructed to obtain the fusion protein. The antigenicity of the protein was analyzed by Western blot. The Lipase activity of the fusion protein was detected by esterase reaction. The location of the protein was tested by the indirect immunofluorescence. The transcription of lipase in different stages of H. contortus was detected by real-time quantification PCR. The results showed that the gene fragment encoding Lipase (without signal peptide region) sized as 864 bp. The recombinant protein weighted as 50 kD was fusion expressed. The fusion protein had highest lipase enzyme activity under 37℃ and pH7. Hc-Lipase was localized at outer and inter membrane as well as the intestinal tracts of adult worms.The expression of Hc-Lipase was higher in the male than other stages. It is indicated that Lipase from H. contortus could play important roles in worm development and modulation of the host immunity. Therefore, it is expected that Hc-Lipase could be a new target in prevention and therapy of Haemonchosis.

CLC Number:

S852.731

ZHOU Li-na, HU Meng-juan, SUN Wei, LI Xiang-rui, XU Li-xin, SONG Xiao-kai, YAN Ruo-feng. Gene Cloning and Biological Characteristics of Lipase Gene from Haemonchus contortus[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(3): 580-587.

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Gene Cloning and Biological Characteristics of Lipase Gene from Haemonchus contortus

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