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Characterization of the third SERK gene in pineapple (Ananas comosus) and analysis of its expression and autophosphorylation activity in vitro Genet. Mol. Biol.
Ma,Jun; He,Yehua; Hu,Zhongyi; Xu,Wentian; Xia,Jingxian; Guo,Cuihong; Lin,Shunquan; Chen,Chengjie; Wu,Chenghou; Zhang,Junli.
Two somatic embryogenesis receptor-like kinase genes (identified as AcSERK1 and AcSERK2) have previously been characterized from pineapple (Ananas comosus). In this work, we describe the characterization of a third gene (AcSERK3) in this family. AcSERK3 had all the characteristic domains and shared extensive sequence homology with other plant SERKs. AcSERK3 expression was studied by in situ hybridization and quantitative real-time PCR to analyze its function. Intense in situ hybridization signals were observed only in single competent cells and competent cell clusters; no hybridization signal was detected in the subsequent stages of somatic embryogenesis. AcSERK3 was highly expressed in embryogenic callus compared to other organs, e.g., 20-80 fold more...
Tipo: Info:eu-repo/semantics/article Palavras-chave: AcSERK3; Autophosphorylation activity; Expression analysis; Prokaryotic expression; Somatic embryogenesis.
Ano: 2014 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572014000400009
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Cloning and Characterization of the Gene Encoding 3-hydroxy-3- Methylglutaryl-coenzyme A (HMG-CoA) Reductase from Fritillaria Cirrhosa D. Don BABT
Zhao,Qi; Li,Rui; Chen,Xiao; Yang,Qian; Li,Jian.
ABSTRACT The enzyme 3-hydroxy-3-methylglutaryl-CoA reductase (HMGR; EC1.1.1.34) catalyzes the first committed step of isoprenoids biosynthesis in Mevalonate (MVA) pathway. Here we report for the first time the cloning and characterization of a full-length cDNA encoding HMGR from Fritillaria cirrhosa (FcHMGR), a bulbous medicinal plant. The full-length cDNA of FcHMGR was 2072 base pair (bp), containing a 1680-bp open reading frame. Bioinformatical analyses revealed that FcHMGR had HMG CoA-binding domains and two NADPH binding domains, which are required for HMGR activity. Quantitative real-time PCR (qRT-PCR) analysis revealed that FcHMGR expressed high in mature bulbs. A truncated version of FcHMGR protein lacking the N-terminal 249-bp GC rich area was...
Tipo: Info:eu-repo/semantics/article Palavras-chave: 3-Hydroxy-3-methylglutaryl-CoA reductases; Fritillaria cirrhosa; Molecular cloning; Expression pattern; Prokaryotic expression.
Ano: 2018 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132018000100438
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Cloning and expression analysis of the chitinase gene Ifu-chit2 from Isaria fumosorosea Genet. Mol. Biol.
Meng,Huimin; Wang,Zhangxun; Meng,Xiangyun; Xie,Ling; Huang,Bo.
Entomopathogenic fungi can produce a series of chitinases, some of which function synergistically with proteases and other hydrolytic enzymes to degrade the insect cuticle. In the present study, the chitinase gene Ifu-chit2 from Isaria fumosorosea was investigated. The Ifu-chit2 gene is 1,435-bp long, interrupted by three short introns, and encodes a predicted protein of 423 amino acids with a 22 residue signal peptide. The predicted Ifu-Chit2 protein is highly homologous to Beauveria bassiana chitinase Bbchit2 and belongs to the glycohydrolase family 18. Ifu-Chit2 was expressed in Escherichia coli to verify chitinase activity, and the recombinant enzyme exhibited activity with a colloidal chitin substrate. Furthermore, the expression profiles of Ifu-chit2...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Isaria fumosorosea; Chitinase; Prokaryotic expression; Quantitative real-time PCR.
Ano: 2015 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572015000300381
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Distribution of Ifitm3 in Yellow-Feathered Broilers and Inhibition of Avian Reovirus Multiplication by Ifitm3 Rev. Bras. Ciênc. Avic.
Wang,Q; Yuan,X; Li,C; Deng,H; Wang,C.
ABSTRACT This study was carried to express the interferon-induced transmembrane protein 3 (IFITM3) in vitro and examine its function in inhibition of avian reovirus (ARV) replication. The recombinant prokaryotic vector expressing yellow-feathered broiler IFITM3 was successfully constructed, and the recombinant protein was expressed in competent Escherichia coli BL21 cells. New Zealand white rabbits were immunized with the purified recombinant protein to prepare a polyclonal antibody, with a titer of 1:128,000. Immunohistochemistry, reverse transcription-PCR, and real-time fluorescence quantitative PCR showed that IFITM3 was distributed in the yellow-feathered broiler immune organs, and the expression of IFITM3 in bursa of Fabricius was more than in spleen...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Yellow-feathered broiler; Interferon; Interferon-induced transmembrane protein 3; Gene cloning; Prokaryotic expression; Avian Reovirus.
Ano: 2018 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-635X2018000200377
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Hyperexpression of two Aspergillus niger xylanase genes in Escherichia coli and characterization of the gene products BJM
Yi,Xiuli; Shi,Yan; Xu,Hui; Li,Wei; Xie,Jie; Yu,Rongqing; Zhu,Jun; Cao,Yi; Qiao,Dairong.
The analysis of individual gene product should enable to clarify the role of a particular enzyme in a complex xylanase system of A. niger. The two genes encoding precursors of co-produced endo-1,4-β-D-xylanases, xynA1 and xynB, were isolated from Aspergillus niger SCTCC 400264 (SCTCC, China) by using RT-PCR technique and then successfully expressed in Escherichia coli BL21. The nucleotide sequences of the xynA1 and xynB genes revealed that they were only 52.5% homology to each other. Characterization of the recombinant enzymes revealed the different properties: the specific activity of recombinant XYNA1 was 16.58 U/mg compared to 1201.7 U/mg for recombinant XYNB; The optimum temperature and pH of the recombinant XYNA1 were 35 ºC and 3.0, respectively,...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Aspergillus niger; Xylanase; Prokaryotic expression; Enzymatic characterization.
Ano: 2010 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822010000300030
Registros recuperados: 5
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