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Cloning and Expression of Ferritin in an Invertebrate Abalone Haliotis Diversicolor Supertexta as Well as Its Modulation by Reactive Species

Author: CaoXiaoHua
Tutor: WuXinZhong
School: Zhejiang University
Course: Aquaculture
Keywords: Abalone abalone Expression of ferritin LPS Active derivative Respiratory burst Immune response Iron effect element Iron regulatory protein
CLC: S917.4
Type: Master's thesis
Year: 2009
Downloads: 17
Quote: 0
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Abstract


Ferritin is the major iron storage protein in everything vivo , has an important role in a number of physiological and pathological processes . This article was successfully cloned abalone abalone ferritin length cDNA sequence . The results are as follows : (1) the cDNA sequence of this 879bp containing a 516bp reading frame encoding 171 amino acids . That the protein has the characteristic of the ferritin structure . Amino acid sequence and invertebrate Haliotis discus hannai homology of up to 97.07% , the Hepu mother of pearl , California sea hare , Crassostrea gigas were 81.3% , 80.7% and 78.9% , respectively ; ( 2 ) semi-quantitative and quantitative RT-PCR showed that ferritin in healthy abalone tissue ( digestive gland , mantle , gills , foot muscle ) were expressed in the digestive gland expression was significantly higher than that of other organizations . LPS stimulation after ferritin gene expression levels and showed significantly increased gradually with time and increased expression levels reached a peak when the stimulus 24h , there is a significant time-dependent , these results table Mingbao ferritin involved in LPS-stimulated immune response; (3) by constructing a recombinant expression plasmid pET -FER , using E. coli BL21 ferritin induced expression and protein purification and preparation of polyclonal antibodies , the results show that : the iron proteins in E. coli highly expressed prepared rabbit polyclonal antibody titer of 1:14000 ; (4) in view of the regulation of ferritin influenced by iron , oxidative stress , biological messenger molecules , cytokines , hormones , etc. , in order to investigate the invertebrate animal respiratory burst in ferritin by regulatory role of ROS and RNS cultivated abalone blood lymphocytes stimulated by LPS -induced respiratory burst to produce large amounts of ROS and RNS the results show that hydrogen peroxide and nitric oxide : stimulation showed significant the increase , and H202 in the regulation of ferritin play a leading role , and can inhibit the expression of ferritin , NO cation resistant to hydrogen peroxide -mediated reduction of ferritin .

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CLC: > Agricultural Sciences > Aquaculture, fisheries > Aquatic basic science > Aquatic Biology > Aquatic Zoology
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