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Y. Li S. L. Yang Z. L. Tang W. T. Cui Y. L. Mu M. X. Chu S. H. Zhao Z. F. Wu K. Li K. M. Peng 《Molecular biology reports》2010,37(1):579-585
As a kind of E3 ligase, the product of FBXL4 gene belongs to a member of FBLs which is the biggest eukaryotic subfamily of F-BOX proteins, it can recognize some substrate
through particular protein–protein interaction domains. To investigate its functions, the polymorphism and association analysis
was analyzed. The partial cDNA of porcine FBXL4 with 2384 bp long was first cloned; the deduced protein comprises a conserved F-BOX domain at position from the 277th to
332nd amino acid. The phylogenetic tree indicated porcine FBXL4 has the closest genetic relationship with bovine FBXL4 than other selected animal species. Ten tissue expression level of porcine FBXL4 mRNA fluctuated remarkably in a large range by quantitative RT-PCR analysis. For two identified SNPs, the genotyping analysis
of Tail showed TT genotype owned dominance in introduced Landrace pig and miniature Guizhou and Wuzhishan breeds, but CC genotype
was more than two other genotypes in miniature Laiwu breed. While in another genotyping analysis of BsaJI, CC genotype was
obviously more than other genotypes in two kinds of Chinese miniature pig breeds and introduced Landrace pig breeds. Furthermore,
the association analysis with immune traits and blood parameters revealed that SNP Tail was significantly associated with
the lymphocyte percentage (P = 0.0166) and the antibody levels for pseudorabies virus vaccination (P = 0.0001) of neonate piglets at 0 day. Meanwhile, SNP BsaJI was significantly associated with lymphocyte percentage of individuals
at 32 days (P = 0.0351), neutrophil percentage (P = 0.0005), the absolute lymphocyte count (P = 0.0458), and the mixed cells (P = 0.0010) of neonate piglets at 0 day. 相似文献
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The extracellular lipase structural gene was isolated from cDNA of Aureobasidium pullulans HN2-3 by using SMARTTM RACE cDNA amplification kit. The gene had an open reading frame of 1245 bp long encoding a lipase. The coding region of the
gene was interrupted by only one intron (55 bp). It encodes 414 amino acid residues of a protein with a putative signal peptide
of 26 amino acids. The protein sequence deduced from the extracellular lipase structural gene contained the lipase consensus
sequence (G-X-S-X-G) and three conserved putative N-glycosylation sites. According to the phylogenetic tree of the lipases,
the lipase from A. pullulans was closely related to that from Aspergillus fumigatus (XP_750543) and Neosartorya fischeri (XP_001257768) and the identities were 50% and 52%, respectively. The mature peptide encoding cDNA was subcloned into pET-24a
(+) expression vector. The recombinant plasmid was expressed in Escherichia coli BL21(DE3). The expressed fusion protein was analyzed by SDS-PAGE and western blotting and a specific band with molecular
mass of about 47 kDa was found. Enzyme activity assay verified the recombinant protein as a lipase. A maximum activity of
0.96 U/mg was obtained from cellular extract of E. coli BL21(DE3) harboring pET-24a(+)LIP1. Optimal pH and temperature of the crude recombinant lipase were 8.0 and 35 °C, respectively and the crude recombinant lipase
had the highest hydrolytic activity towards peanut oil. 相似文献
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Yanyang Liu Junzhou Li Yuling Li Mengguan Wei Qingxin Cui Qilei Wang 《Molecular biology reports》2010,37(2):755-761
A full-length cDNA encoding a maize GTP-binding protein of the ADP-ribosylation factor family was cloned by suppression subtractive
hybridization and an in silico cloning approach. The cDNA was 938 bp in length and contained a complete ORF of 612 bp, which
encodes a protein of 203 amino acid residues. Its deduced amino acids sequence had an 83% identity with that of a GTP-binding
protein in rice. The gene was designated ZmArf2. The ZmArf2 gene consists of G1, G2, G3, G4 and G5 boxes, and Switch I and Switch II regions. Eight nucleotides differed and five amino
acids changed between the popcorn inbred N04 and the dent corn inbred Dan232. One changed amino acid was in the G1 box. RT-PCR
analysis showed that ZmArf2 expression increased in the early stages of endosperm development and was not tissue-specific. 相似文献
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The RT PCR and RACE methods were used to obtain the cDNA sequence of an APX gene of muskmelon after the leaves were induced
with powdery mildew. The cDNA length of the APX gene is 1,047 bp with a 750 bp ORF encoded a 249 amino acid and the molecular
weight of APX protein is 27.3 kDa. The analysis showed that the CmAPX genomic DNA contained 10 extrons and 9 introns. The identity of the amino acid sequence deduced from the cDNA with the
APX family of other homologous members was about 74–97%. A Full-length of ORF was sub-cloned into prokaryotic expression vector
pET24a. The recombinant proteins had high expression level in E. coli. Analysis of expression at mRNA level showed that CmAPX exhibited highly tissue-specific patterns of expression. The mRNA level and enzyme activities assays showed that CmAPX might play an important role in the pathogenesis of powdery mildew. 相似文献
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Biomacromolecules import into the nucleus is a complex progress which requires the participation of several cytosolic factors,
and nuclear transport factor 2 (NTF2) is one of essential components in nuclear trafficking. Its main role is to transport
RanGDP from cytoplasm to nucleus by interacting with FxFG nucleoporin repeats. In the study a putative new gene, designated
as CcNTF2, was obtained from the moss (Conocephalum conicum) cDNA library we have constructed. The full-length cDNA sequence is 913 bp in size contains a 372 bp open reading frame (ORF)
flanked by a 195 bp 5′-untranslated sequence and a long 346 bp 3′-non-coding region, encoding 123 amino acids of 13,575.3 Da.
Part of the genomic sequence was also cloned and sequenced, which is 1,602 bp long and possesses two exons and one intron.
Alignment analysis showed that the CcNTF2 protein is high conserved among plant NTF2 and shares 81% similarity with the ones
from Arabidopsis thaliana and Brassica rapa. The expression of wild-type CcNTF2 was detected by immunoblotting of extraction of C. conicum and it indicated the putative protein is integral. Through functional expression of CcNTF2-green fluorescent protein (GFP)
in tobacco, it was demonstrated that CcNTF2 can accumulate at the nuclear rim. Site-directed mutagenesis analysis confirmed
CcNTF2 P71K has influence on the protein import into nucleus. In addition, overexpression of CcNTF2 P71K was observed to be
deleterious for the plant cell. It is the first illumination of NTF2 in moss, and our study established the primary foundation
for further research on moss NTF2. 相似文献
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Zhisheng Zhang Delin Mo Peiqing Cong Zuyong He Fei Ling Anning Li Yuna Niu Xiao Zhao Chunyan Zhou Yaosheng Chen 《Molecular biology reports》2010,37(3):1611-1618
The product of transmembrane and coiled-coil domains 1 (TMCO1) gene is a member of DUF841 superfamily of several eukaryotic proteins with unknown function. The partial DNA sequence of
porcine TMCO1 was first cloned with a pig 567 bp ORF encoding 188 amino acids. By tissues expression analysis, the TMCO1 was found highly expressed in the liver, kidney and heart. The porcine TMCO1 protein was subsequently demonstrated to localize
in the mitochondrion by confocal fluorescence microscopy. This data provides an important basis for conducing further studies
on the functions and regulatory mechanisms underlying the role of TMCO1 gene. 相似文献
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Lijun Chai Manosh Kumar Biswas Xiaoxia Ge Xiuxin Deng 《Plant Molecular Biology Reporter》2010,28(4):569-577
Plant s-phase kinase-associated protein 1 (SKP1) genes have diverse functions in plant developmental and physiological activities. Herein, we described a novel SKP1 gene, designated as CgSKP1, from ‘Shatian’ pummelo (Citrus grandis Osbeck). The cDNA sequence of CgSKP1 was 603 bp and contained an open reading frame of 477 bp. Genomic sequence of the CgSKP1 gene contained two exons and one intron. The predicted amino acid sequence of this gene is consisted of 158 amino acids with
theoretical proteins size of 17.9 kDa. CgSKP1 had high identity with SKP1 genes from other plant species within two conserved region. Full-length cDNAs were also amplified and cloned from six citrus
varieties, with 95% nucleotide identity and about 98% amino acid similarity among them. Gel blot analysis suggested that CgSKP1 existed as a single locus in the ‘Shatian’ pummelo genome. The expression of CgSKP1 was gradually increased during flower developmental stages in ‘Shatian’ pummelo. Moreover, expression analysis by RT-PCR,
qRT-PCR and in situ hybridization of CgSKP1 showed that it was highly expressed in the leaf, petal, anther and ovary, but lowly in the style. These findings indicated
that CgSKP1 was closely related to ‘Shatian’ pummelo flower development. 相似文献
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Jian-Xia Zhang Kun-Lin Wu Li-Ning Tian Song-Jun Zeng Jun Duan 《Acta Physiologiae Plantarum》2011,33(2):409-417
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The cell divisions cycle 42 (Cdc42) gene has been characterized in the fungi, such as Candida albicans, Penicillium marneffei, and Wangiella (Exophiala) dermatitidis, which plays important roles during growth and development. The partial cDNA sequence of Cdc42 of Fonsecaea monophora (F. monophora), designated FmCdc42, was obtained using degenerate primers based on the conserved domain of the other fungi Cdc42. Then the complete cDNA sequence of FmCdc42 was obtained by 5′ and 3′ RACE. The full-length cDNA is 1,510 bp in size which had an open reading frame (ORF) of 582 bp,
encoding 193 amino acid residues. The predicted molecular mass of FmCdc42 is 21.5 kDa with an estimated theoretical isoelectric point of 5.67. The deduced amino acid sequence of FmCdc42 shows 99% identity to that of Wangiella (Exophiala) dermatitidis. 5 exons and 4 introns are identified within the 1,617 bp FmCdc42 genomic DNA sequence of F. monophora. The ORF could be subcloned into the pCDNA6/myc-His B expression vector. The recombinant protein about 27.5 kD infusion protein
had high expression level in Vero cells with SDS-PAGE and Western blot analysis. Quantitative real time RT-PCR revealed that
FmCdc42 was the highest expression in the sclerotic bodies’ stage compared with that in the mycelia and conidia stages, which indicated
that the FmCdc42 may be involved in formation of F. monophora sclerotic bodies. 相似文献
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Jing Lan Ming-Gang Lei Yi-Bing Zhang Jian-Hua Wang Xiao-Ting Feng De-Quan Xu Jian-Fang Gui Yuan-Zhu Xiong 《Molecular biology reports》2009,36(7):2003-2010
To investigate the differential expression of genes in the skeletal muscle between Yorkshire and Chinese indigenous breed
Meishan pigs, suppression subtractive hybridization was carried out and many genes were proved to be expressed significantly
different in the two breeds. One gene highly expressed in Meishan but lowly expressed in Yorkshire specific library, shared
strong homology with human pyruvate dehydrogenase kinase 4 (PDK4). Using semi-quantity and quantity PCR, We confirmed its differential expression between the two breeds. Temporal and spatial
expression analysis indicated that porcine PDK4 gene is highly expressed in skeletal muscle and the highest in neonatal pigs. Complete cDNA cloning and sequence analysis
revealed that porcine PDK4 gene contains an open reading frame of 1,221 bp. The deduced amino acid sequence showed conservation in evolution. A G/A
mutation in intron 9 was identified and association analysis showed that it was significantly associated with intramuscular
fat, muscle water content. 相似文献
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The adipose triglyceride lipase (PNPLA2, also known as ATGL) is a novel triacylglycerol (TG) lipase which specifically removes
the first fatty acid from the triglyceride molecule generating free fatty acid and diglyceride (DG) in mammalian cells. Here
we describe the molecular characterization of the porcine ATGL gene. The full-length cDNA sequence contains a 1,461 bp open reading frame encoding a protein of 486 amino acids with a calculated
molecular mass of 53.2 kDa and an isoelectric point of 7.90. The porcine ATGL protein shares high identity with other mammalian
ATGL. The ATGL gene contains 9 coding exons, spans approximately 6 kb. The porcine ATGL mRNA was expressed predominantly in backfat, mildly in muscle, small intestine and heart, and almost absent in liver, spleen,
lung, stomach, kidney and ovary. Statistical analysis showed the ATGL gene polymorphism (G/A392) was different between Chinese indigenous and introduced commercial western pig breeds, and was highly associated with almost
all the fat deposition and carcass traits, including subcutaneous fat thickness, viscera adipose tissue, lean percentage,
loin eye traits and even rib numbers. 相似文献
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A chalcone reductase (CHR) gene was isolated from Astragalus membranaceus Bge. var. mongholicus (Bge.) Hsiao (A. mongholicus). The full-length cDNA of A. mongholicus CHR, designated as Amchr (GenBank accession No. HM357239), was 1196 bp long. It had a 957 bp open reading frame encoding a 318-amino acid protein
of 35 kDa, a 67 bp 5′ non-coding region and a 172 bp 3′-untranslated region. The putative AmCHR protein showed striking similarity
to CHR from other leguminous species. Two-dimensional structure modeling showed that AmCHR consisted of 45.28% α-helix, 10.38%
extended strand and 44.34% random coil. Prediction showed that three-dimensional AmCHR was a global protein containing an
aldo-ket-red domain, with a putative Asp-Tyr-Lys-His catalytic tetrad in the center. The AmCHR gene was 1251 bp long, consisting
of three exons and two introns. Intron I was 125 bp and intron II was 169 bp long. Southern blot analysis indicated that Amchr belonged to a small multigene family. Under natural conditions, Amchr was expressed differentially in the root, stem and leaf tissues of A. mongholicus, with a preferential expression in the root. The recombinant AmCHR protein was successfully expressed in Escherichia coli strain BL21 with pET42a vector. The result showed that the expressed AmCHR protein had molecular weight of about 35 kDa,
which matched the size of the predicted protein by bioinformatic analysis. This study opened avenues towards understanding
of the function of AmCHR protein and the role of the Amchr gene in the calycosin-7-O-β-d-glucoside branch pathway in A. mongholicus. 相似文献
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Jiquan Zhang Yuying Sun Fuhua Li Bingxin Huang Jianhai Xiang 《Molecular biology reports》2010,37(4):1913-1921
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Xuewen Xu Haifang Qiu Zhi-Qiang Du Bin Fan Max F. Rothschild Fan Yuan Bang Liu 《Molecular biology reports》2010,37(1):451-459
CRP3 is the muscle-specific form of the cysteine and glycine-rich protein family and plays an important role in myofiber differentiation.
Here we isolated and characterized its coding gene CSRP3 from porcine muscle. Phylogenic analyses demonstrated that CSRP3 diverged first and is distinguished from two other members, CSRP1 and CSRP2. CSRP3 mRNA was up-regulated during the development of porcine embryonic skeletal muscle, indicating its potential importance in
muscle growth. Genetic variant analyses detected multiple variations in an approximately 400 bp region covering exon 4 and
its downstream intron, and two haplotypes were identified by sequencing. One of synonymous substitutions C1924T was used for
linkage and association analyses. It was revealed that the substitution of C1924T had significant associations with firmness
(P < 0.01), Lab Loin pH, Off Flavor Score and Water Holding Capacity (P < 0.05), and a suggestive effect (P < 0.1) on Flavor Score and Average Glycolytic Potential in a Berkshire × Yorkshire F2 population. The association analyses
results agreed with the gene’s localization to a QTL region for meat quality traits on porcine chromosome 2p14-17 demonstrated
by both linkage mapping and RH mapping. These results provide fundamental evidence for CSRP3 as a functional candidate gene affecting pig meat quality. 相似文献