Microplusin-like Gene And Its Expression Analysis Of Haemaphysalis Flava

Microplusin is an important component of tick defense system,which has many members and high content.These proteins can inhibit a variety of pathogenic microorganisms,and can be developed as new antimicrobial agents for livestock and poultry disease prevention or feed preservation.In this paper,we first collected Micropulsins from Haemaphysalis flavas and analyzed the structural characteristics of some of them.Then,we cloned the full-length genes of Hfmicropulsin-1 and Hfmicropulsin-2 by RACE,and analyzed the expression levels of the two genes in different developmental stages,different degree of blood saturation and different tissue parts by q PCR,The purpose of this study is to lay a foundation for exploring the physiological role of Microplusin in ticks.In order to understand the structural characteristics of tick like Microplusin protein and the sequence structure and expression of some members,we first searched the ovarian transcriptome Library of Haemaphysalis flavas with Microplusin as the key word,and obtained 13 complete sequences for online analysis.The results showed that the Micropulsin length of H.flavas was 99-115 aa;Its N-terminal sequence is "MKA",and its C-terminal is rich in histidine,with six "C" residues scattered between them;It has signal peptide.The alignment showed that they were divided into six groups.The sequences within the groups were very similar,the tertiary structure was the same,and the sequences between groups were quite different.The tertiary structure was significantly different in four regions.Then,the two terminal deletion sequences of cluster-19051.0 and cluster-24081.34005 were amplified by RACE,and the full length c DNA of two Microplusins,named Hfmicroplusin-1 and Hfmicroplusin-2,were obtained by splicing.The full-length of Hfmicroplusin-1 is 581 bp,the ORF frame size is 309 bp,encoding 102 aa protein.The full-length of Hfmicroplusin-2 is 497 bp,the ORF frame size is 309 bp,encoding 102 aa protein.Both of them have signal peptides and have some differences in protein structure.At the same time,both of them have antibacterial effect.Finally,RT-PCR was used to detect the expression levels of Hfmicroplusin-1 and Hfmicroplusin-2 genes in different developmental stages,different tissue parts and different degree of fullness of H.flavas.The results showed that Hfmicroplusin-1 and Hfmicroplusin-2 genes were expressed in young,young and adult stages,but the highest expression was found in female adult;The highest expression level was found in 1 / 4 of full blood;The highest expression level was found in salivary gland,body shell and ovary.