|Table of Contents|

Cloning and Construction of VIGS Expression Vector of LtXTH5 in Lilium tsingtauense

《北方园艺》[ISSN:1001-0009/CN:23-1247/S]

Issue:
2020年15
Page:
74-81
Research Field:
Publishing date:

Info

Title:
Cloning and Construction of VIGS Expression Vector of LtXTH5 in Lilium tsingtauense
Author(s):
CHI Xiufeng; JIANG Xinqiang; LI Yanshuo; LI Wei; WANG Kuiling; LIU Qinghua
(College of Landscape Architecture and Forestry，Qingdao Agricultural University，Qingdao，Shandong 266109)
Keywords:
Lilium tsingtauense; LtXTH5; cloning; sequence analysis; silence vector
PACS:
-
DOI:
10.11937/bfyy.20193398
Abstract:
Using the tepals of Lilium tsingtaense as the test material，LtXTH5 gene was isolated from Lilium tsingtaense by RT-PCR，and the bioinformatics analysis and the construction of virus induced silencing expression vector were carried out.The basic physical and chemical properties and vector construction method of the flower development gene LtXTH5 were studied in order to provide the basis for the verification and identification of biological functions of LtXTH5 in the later stage.The results showed that the open reading frame of LtXTH5 gene (MN245449) contained 882 bp and encoded 293 amino acids.Bioinformatics analysis predicted that the relative molecular weight of LtXTH5 was 33.8 kDa，the molecular formula was C1536H2283N407O436S12，the theoretical isoelectric point (pI) was 7.03，the fat index was 68.23，and the instability index was 31.68.LtXTH5 was a hydrophilic protein with signal peptide.Motif analysis of XTHs from different plants showed that the number of conserved motif was different among different plants.LtXTH5 contains nine motifs.Sequence alignment indicated LtXTH5 was a typical member of XTH family，which include DEIDFEFLG catalytic active sites and GH16_XET domain.Phylogenetic analysis showed that LtXTH5 was closely related to AtXTH5 in Arabidopsis thaliana，belonging to I/II subfamily，and had transglycosylase (XET) or hydrolase (XEH) activities.Using the 5′ end 537 bp fragment of LtXTH5 and pTRV2 vector plasmid，a silencing vector (TRV2-LtXTH5) was successfully constructed，and TRV2-LtXTH5 vector was successfully transferred to agrobacterium GV3101.In the next step，we plan to use the agrobacterium liquid which had been transferred into TRV2-LtXTH5 vector to infect tobacco and further explore the function of LtXTH5 gene.

References:

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Last Update: 2020-10-28