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1.
In the present study, a ras-like nuclear (Ran) gene was obtained from the ovary and neurosecretory organ in eyestalk cDNA
library of black tiger prawn (Penaeus monodon). The full-length black tiger prawn Ran (PmRan) cDNA consisted of 1140 nucleotides including an open reading frame (ORF) 648 bp, a 5′ untranslated region (5′UTR) of 117 bp
and a 3′UTR of 375 bp with a polyadenylation signal sequence “aataaa” and a poly (A) tail. The ORF encoded a peptide of 215
amino acids with molecular mass 24.6 kDa and a theoretical isoelectric point of 7.39. ScanProsite analysis indicated that
PmRan protein sequence contained a small GTPase Ran family motif. Homology analysis of the deduced amino acid sequence of the PmRan with other known Ran sequences by MatGAT software revealed that the PmRan show very high homology with the sequences of other animals (92.1–98.6% similarity, 85.6–98.1% identity). Analysis of the
tissue expression pattern of the PmRan gene showed that the PmRan mRNA was expressed in all tested tissues, including hepatopancreas, ovary, muscle, intestine, neurosecretory organ in eyestalk,
neurosecretory organ in brain, stomach, and heart, with the highest levels in ovary. Furthermore, the PmRan expression was found to be high level in the six ovarian stages of development. The results indicated PmRan might play an important role in ovarian development. 相似文献
2.
The techniques of homology cloning and anchored PCR were used to clone the fucose-binding lectin (F-type lectin) gene from
Japanese sea perch (Lateolabrax Japonicus). The full-length cDNA of sea perch F-lectin (JspFL) contained a 5′ untranslated region (UTR) of 39 bp, an ORF of 933 bp
encoding a polypeptide of 310 amino acids with an estimated molecular mass of 10.82 kDa and a 3′ UTR of 332 bp. The searches
for nucleotides and protein sequence similarities with BLAST analysis indicated that the deduced amino acid sequence of JspFL
was homological to the Fucose-binding lectin in other fish species. In the JspFL deduced amino acid sequence, two tandem domains
that exhibit the eel carbohydrate-recognition sequence motif were found. The temporal expressions of gene in the different
tissues were measured by real-time PCR. And the mRNA expressions of the gene were constitutively expressed in tissues including
spleen, head-kidney, liver, gill, and heart. The JspFL expression in spleen was different during the stimulated time point,
2 h later the expression level became up-regulated, and 6 h later the expression level became down-regulated. The result indicated
that JspFL was constitutive and inducible expressed and could play a critical role in the host-pathogen interaction. 相似文献
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4.
The products of mammalian LPIN2 and LPIN3 are phosphatidate phosphatase type 1 enzymes, which play an important role in the de novo biosynthesis of triacylglycerol,
phosphatidylcholine and phosphatidylethanolamine. In this study, we obtained a 2,985-bp cDNA sequence of porcine LPIN2, which contains a 2,676-bp open reading frame flanked by an 11-bp 5′UTR and a 298-bp 3′UTR, and a 2,843-bp cDNA sequence
of porcine LPIN3, which contains a 111-bp 5′UTR, a 2,580-bp open reading frame and a 152-bp 3′UTR. RT-PCR analysis showed that both LPIN2 and LPIN3 mRNA were ubiquitously expressed with a very high level in liver. By using the somatic cell hybrid panel (SCHP) and the radiation
hybrid (IMpRH) panel, porcine LPIN2 and LPIN3 were assigned to 6q24-(1/2)q31 and 17(1/2)q21-q23, respectively. One T2193C single nucleotide polymorphism in LPIN2 was identified and was detected by Hin6I PCR-RFLP. Association analysis showed that different genotypes of LPIN2 were associated with back-fat thickness between the 6th and 7th ribs (P < 0.01). 相似文献
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He XP Xu XW Zhao SH Fan B Yu M Zhu MJ Li CC Peng ZZ Liu B 《Molecular biology reports》2009,36(5):1175-1180
Lpin1 deficiency prevents normal adipose tissue development and remarkably reduces adipose tissue mass, while overexpression of
the Lpin1 gene in either skeletal muscle or adipose tissue promotes adiposity in mice. However, little is known about the porcine Lpin1 gene. In the present study, a 5,559-bp cDNA sequence of the porcine Lpin1 gene was obtained by RT-PCR and 3′RACE. The sequence consisted of a 111-bp 5′UTR, a 2,685-bp open reading frame encoding
a protein of 894 amino acids and a 2,763-bp 3′UTR. Semi-quantitative RT-PCR analysis revealed that Lpin1 had a high level of expression in the liver, spleen, skeletal muscle and fat, a low level of expression in the heart, lung
and kidney. The porcine Lpin1 gene was assigned to 3q21-27 by using the somatic cell hybrid panel (SCHP) and the radiation hybrid (IMpRH) panel. One C93T
single nucleotide polymorphism (SNP) was identified and genotyped using the TaqI PCR-RFLP method. Association analysis between the genotypes and fat deposition traits suggested that different genotypes
of the Lpin1 gene were associated with percentage of leaf fat and intramuscular fat. 相似文献
8.
Cloning and expression of stearoyl-CoA desaturase 1 (SCD-1) in the liver of the Sichuan white goose and landes goose responding to overfeeding 总被引:2,自引:0,他引:2
The EST sequence of goose (Anser cygnoides) Stearoyl-CoA desaturase 1(SCD-1) was obtained from a subtractive cDNA library. To further investigate the role of SCD-1
in lipid metabolism in geese, 5′-RACE and 3′-RACE were carried out in this study to obtain the complete cDNA sequence of goose
SCD-1, which contained a 29-bp 5′ UTR, a 1074-bp open reading frame (ORF) encoding 357 amino acids, and a 125-bp 3′ UTR. The
expression of SCD-1 was measured in several tissues, and the effects of overfeeding on the expression of SCD-1 were studied.
The results of real time RT-PCR demonstrated that, compared to the brain, goose SCD-1 mRNA was more abundant in the liver.
Overfeeding markedly increased the mRNA expression of SCD-1 in the liver of Sichuan White and Landes geese, and gene expression
was markedly higher in the Sichuan White goose than in the landes goose. The mRNA abundance of SCD-1 in the liver had significant
positive correlations with triacylglycerol (TG) content in liver lipids and in the levels of plasma insulin and very low-density
lipoproteins (VLDL) levels in Sichuan white geese. However, the mRNA abundance of SCD-1 in the livers of Landes geese had
only significant positive correlations with the TG content in liver lipids. In conclusion, SCD-1 is not only critical for
hepatic steatosis in geese but is also important for the difference in lipid deposition in the livers of the two breeds. 相似文献
9.
The techniques of homology cloning and anchored PCR were used to clone the Hsp90 gene from black tiger shrimp. The full length
cDNA of black tiger shrimp Hsp90 (btsHsp90) contained a 5′ untranslated region (UTR) of 72 bp, an ORF (open reading frame)
of 2160 bp encoding a polypeptide of 720 amino acids with an estimated molecular mass of 83-kDa and a 3′ UTR of 288 bp. The
sequence of the coding region showed 90 and 84% homology with that of the Chiromantes haematocheir and Homo sapiens, respectively. Conserved signature sequences of Hsp90 gene family were found in the btsHsp90 deduced amino acid sequence.
The temporal expressions of Hsp90 gene were constitutively in the black tiger shrimp tissues including liver, ovary, muscle,
brain stomach, and heart, and their levels were markedly enhanced after 30-min heat treatment at 37°C. In ovarian maturation
stages, the expression of btsHsp90 was strongest in the second stage, weaker in the fourth and first stage. 相似文献
10.
Heat shock protein 90 (Hsp90), known as molecular chaperone, is involved in protein folding and assembly in the cell. In the
present study, a full-length cDNA named Vitis pseduoreticulta heat shock protein 90 (VpHsp90) (GenBank accession Number:EU239815), encoding a heat shock protein 90, was obtained by degenerated primers and 3′-and 5′-RACE
from Vitis pseudoreticulata according to our previously obtained EST sequence (GenBnak accession number:DV182112), putatively known as Hsp90. Comparison of VpHsp90 sequence has revealed that an open reading frame (ORF) consists of 2,100 bp nucleotides and the translated proteins of 699
amino acid residues. The molecular mass of VpHsp90 calculated from the deduced amino acid sequence was 80.2 kDa, Isolectric Point was 4.893, which is in close proximity of
Hsp90. The maximum similarity of VpHsp90 at nucleotides level (85%) and protein level (96%) was found to be with Nicotiana tabacum. Phylogenetic tree analysis at both the nucleotides and amino acids levels indicates that Vitis pseduoreticulata, Nicotiana tabacum, and Arabidopsis thaliana Hsp90 sequences comprise one clade, which is closely related to Oryza sativa, Hordeum vulgare and Triticum aestivum Hsp90s. It may be reasonably concluded that VpHsp90 possesses the ancestral gene of Hsp90 similar to that of higher plant species. 相似文献
11.
Molecular cloning and expression analysis of a cytosolic Hsp70 gene from Antarctic ice algae Chlamydomonas sp. ICE-L 总被引:1,自引:0,他引:1
Shenghao Liu Pengying Zhang Bailin Cong Chenlin Liu Xuezheng Lin Jihong Shen Xiaohang Huang 《Extremophiles : life under extreme conditions》2010,14(3):329-337
A cDNA encoding heat shock protein 70 of Antarctic ice algae Chlamydomonas sp. ICE-L (designated as CiHsp70) was identified by RT-PCR and rapid amplification of cDNA ends approaches. The full-length
cDNA of CiHsp70 was 2,232 bp, consisting of a 5′-terminal untranslated region (UTR) of 76 bp, a 3′-terminal UTR of 203 bp
with a poly (A) tail, and an open reading frame of 1,953 bp. The CiHsp70 cDNA encoded a polypeptide of 651 amino acids with
an ATPase domain of 388 amino acids, the substrate peptide binding domain of 246 amino acids and a C-terminus domain of 17
amino acids. The inducible CiHsp70 cDNA was highly homologous to other plant cytosolic Hsp70 genes and clustered together
with green algae and higher plant rather than brown algae, diatom and Cryptophyta. Antarctic ice algae were treated with different
stress conditions and messenger RNA (mRNA) expression levels of CiHsp70 were quantified by quantitative RT-PCR. The results
showed that both cold and heat shock treatments could stimulate CiHsp70 mRNA expression. Meanwhile, CiHsp70 mRNA expression
level increased 2.9-fold in response to UV-B radiation for 6 h, while the expression levels of CiHsp70 were remarkably increased
after removing the UV-B radiation and immediately providing additional 6 h visible light. Furthermore, treating with 62 or
93‰ NaCl for 2 h, CiHsp70 mRNA expression level increased 3.0- and 2.1-fold, respectively. Together, our observations revealed
that CiHsp70 as a molecular chaperone might play an important role in Antarctic ice algae Chlamydomonas sp. ICE-L acclimatizing to polar environment. 相似文献
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FaLin Zhou ShiGui Jiang JianHua Huang Lihua Qiu Dianchang Zhang Tiannfeng Su 《Molecular biology reports》2011,38(3):1921-1927
In present study, a QM gene was obtained from the ovary and neurosecretory organ in eyestalk cDNA library of black tiger prawn
(Penaeus monodon). The full-length black tiger prawn QM (PmQM) cDNA contained a 5′-UTR of 41 bp, an ORF of 663 bp encoding a polypeptide of
220 amino acids with molecular weight 25.5 kDa, and a 3′-UTR of 54 bp. Homology analysis of the deduced amino acid sequence
of the PmQM with other known QM sequences by MatGAT software revealed that the PmQM was high homology with other invertebrates.
A conserved signature sequence of the QM family was found in the PmQM deduced amino acid sequence. Analysis of the tissue
expression pattern of the PmQM gene showed that the PmQM mRNA was expressed in all tissues tested, with highest levels in
ovary. Furthermore, the PmQM expression was found to be different in three important ovarian stages of development. The results
indicated PmQM might play an important role in ovarian development. 相似文献
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Yubero N Esteso G Cardona H Morera L Garrido JJ Barbancho M 《Molecular biology reports》2009,36(7):1957-1965
This work describes the cloning and structural analysis of a Tpt1 cDNA coding for the porcine translationally controlled tumor protein (TCTP) molecule and its expression in porcine cells
and tissues. Pig Tpt1 cDNA is 842-pb long that displays typical features of translationally controlled mRNAs, including a 5′-UTR containing a 5′-terminal
oligopyrimidine tract (5′-TOP), and a 3′-UTR with a high CG-content and one AU rich element (ARE). Both 5′-UTR and 3′-UTR
are highly conserved when they are compared with those of other mammals. The pig Tpt1 cDNA contains a 516-b open reading frame that encodes a predicted TCTP protein composed of 172 amino acids that exhibits
extensive conservation compared with TCTP sequences from other species and a common structural feature with all the other
TCTP proteins analyzed in mammals. Expression analysis demonstrated that Tpt1 mRNA is ubiquitously expressed in normal porcine tissues and cells, showing a higher expression in spleen, lymph nodes and
lung, and a lower one in skin and heart. The pig Tpt1 gene localizes on the porcine chromosome 11, region p11. 相似文献
16.
Expression Enhancement of a Rice Polyubiquitin Gene Promoter 总被引:11,自引:0,他引:11
An 808 bp promoter from a rice polyubiquitin gene, rubi3, has been isolated. The rubi3 gene contained an open reading frame of 1140 bp encoding a pentameric polyubiquitin arranged as five tandem, head-to-tail
repeats of 76 aa. The 1140 bp 5′ UTR intron of the gene enhanced its promoter activity in transient expression assays by 20-fold.
Translational fusion of the GUS reporter gene to the coding sequence of the ubiquitin monomer enhanced GUS enzyme activity in transient expression assays
by 4.3-fold over the construct containing the original rubi3 promoter (including the 5′ UTR intron) construct. The enhancing effect residing in the ubiquitin monomer coding sequence
has been narrowed down to the first 9 nt coding for the first three amino acid residues of the ubiquitin protein. Mutagenesis
at the third nucleotide of this 9 nt sequence still maintains the enhancing effect, but leads to translation of the native
GUS protein rather than a fusion protein. The resultant 5′ regulatory sequence, consisting of the rubi3 promoter, 5′ UTR exon and intron, and the mutated first 9 nt coding sequence, has an activity nearly 90-fold greater than
the rubi3 promoter only (without the 5′ UTR intron), and 2.2-fold greater than the maize Ubi1 gene promoter (including its 5′ UTR intron). The newly created expression vector is expected to enhance transgene expression
in monocot plants. Considering the high conservation of the polyubiquitin gene structure in higher plants, the observed enhancement
in gene expression may apply to 5′ regulatory sequences of other plant polyubiquitin genes. 相似文献
17.
Junguang He Zhigang Dong Zhiwei Jia Jianhua Wang Guoying Wang 《Molecular biology reports》2010,37(2):865-874
18.
The full length of major histocompatibility complex (MHC) class IIB cDNA was cloned from a Chinese population of Paralichthys olivaceus by homology cloning and rapid amplification of cDNA ends-polymerase chain reaction (RACE-PCR). The MHC IIB genomic sequence
is 1,864 bp long and consists of 34-bp 5′UTR, 741-bp open reading frame, 407-bp 3′UTR, 96-bp intron1, 392-bp intron2, 85-bp
intron3, and 109-bp intron4. Phylogenetic analysis showed that the putative MHC class IIB amino acid of the Chinese P. olivaceus shared 28.3% to 85.4% identity with that of the reported MHC class IIB in other species. A significant association between
MHC IIB polymorphism and disease resistance/susceptibility was found in Chinese P. olivaceus. Thirteen different MHC IIB alleles were identified among 411 clones from 84 individuals. Among the 280 (268) nucleotides,
32 (11.4%) nucleotide positions were variable. Most alleles such as alleles a, b, c, d, e, f, j, k, i, m were commonly found in both resistant and susceptible stock. Via χ2 test, allele d was significantly more prevalent in individuals from susceptible stock than from resistant stock, and their percentages were
23.80% and 7.14%, respectively. In addition, allele g occurred in 9 and allele h in 4 of 42 resistant individuals that were not present in the susceptible stock; their percentages were 21.4% and 9.52%,
respectively. Although allele l was found only in 8 individuals from the susceptible stock, its percentage is 19.05%. 相似文献
19.
Babiker HA Nakatsu Y Yamada K Yoneda A Takada A Ueda J Hata H Watanabe T 《Immunogenetics》2007,59(1):59-67
Millennia-long selective pressure of single-strand RNA viruses on the bovine Mx locus has increased the advantages of using the bovine Mx protein to evaluate the ultimate significance of the antiviral
role of Mx proteins. The conclusions of research based only on the bovine Mx1 protein showed the need for comprehensive studies
that demonstrate the role of all isoforms, individually or together, especially in the presence of a second isoform, the bovine
Mx2 gene. This study provides information about bovine and water buffalo Mx2 genes, as well as their allelic polymorphism and basic antiviral potential. Observation of an Mx2 cDNA sequence (2,381 bp) obtained from 15 animals from 11 breeds using primers based on a previous sequence (NCBI accession
no. AF335147) revealed several nucleotide substitutions, with eight different alleles and two amino acid exchanges: Gly to
Ser at position 302 and Ile to Val at position 354, though the latter was found only in the NCBI database. A water buffalo
Mx2 cDNA sequence was identified for the first time, revealing 46 nucleotide substitutions with 12 amino acid variations, in
addition to a 9-bp insertion in the 5′ untranslated region UTR, compared with the bovine Mx2 cDNA. Transfected 3T3 cells expressing bovine Mx2 mRNAs coding Gly or Ser at position 302, water buffalo Mx2 mRNA, positive control bovine Mx1 mRNA-expressing cells, and negative control parental 3T3 were subjected to infection with recombinant vesicular stomatitis
virus (VSVΔG*-G), as were empty pCI-neo vector-transfected cells. The positive control and all cells expressing Mx2 mRNAs displayed significantly higher levels of antiviral activity against VSVΔG*-G (P < 0.01) than did the negative controls. 相似文献