Expression Analysis of Apostichopus Japonicus Trypsin-like Serine Protease Gene during

Regeneration
Author :YuanZhe
Tutor:HouLin
School :Liaoning Normal University
CLC :S917.4
TYPE :Masters thesis
Download the PDF Full Text:http://www.topresearch.org/showinfo-177-678067-0.html
Year:2012
Abstract:
Trypsin-like serine protease (TLS) plays an important role in many physiological processes including
wound healing, phlogosis reaction, blood clotting, regeneration etc.In this paper, a1216bp full-length
cDNA sequence of TLS including39bp5UTR and355bp3UTR was cloned from Apostichopus
japonicus by means of the RACE technique for the first time. we obtained the whole DNA sequence
of TLS including intron and exon by means of Genewalking and the result revealed that there
are4introns and5exons in this DNA sequence. In-situ hybridization (ISH) was taken to analyse the
distribution of TLS and the results revealed that the TLS gene was widely distributed in body wall,
intestine and respiratory trees. The signals are strongest in intestine. The expression patterns during
different regeneration stages of the TLS gene in the body wall, intestine and respiratory trees were
investigated using real-time quantitative PCR. The results show that there was a remarkable and
temporary up-regulation of TLS gene expression in the body wall within one hour and subsequent
down-regulation of TLS similar to intestine and respiratory trees. With the recovery of tissues, the
expression level of the TLS gene was gradually up-regulated and finally reached normal levels. But in
body wall, intestine and respiratory trees, the expression level of TLS was lower than the normal
tissues until the regeneration of new tissues. TLS was regulated during different regeneration stages
suggesting that TLS is important in the regeneration process of Apostichopus japonicus. Our research
of Apostichopus japonicus TLS gene expression pattern during different regeneration stages will make
great sense to illustrate relevant molecular mechanism and Regeneration regulatory pathways.
Meanwhile, our research have great application value to maintain quality of Apostichopus japonicus
during aquaculture process. Meanwhile, our research have great application value to further
understand the occurrence of disgorging entrails and improve the survival rate of larval during larval
production and aquaculture of Apostichopus japonicus