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In order to identify the genes associated with glioblastoma differentiation, some ESTs, expressed differentially in the control cell and the differentiated human glioblastoma cell line BT-325 induced by the all-trans retinoid acid, have been isolated by the method of DDRT-PCR. Of the 46 ESTs sequenced, 19 are from new genes. A full-length 1 535-bp cDNA, termed gene GDR1, has been isolated from the human cDNA library using the probe designed according to one of the novel ESTs, HGBB098. The open reading frame of GDR1 gene encodes a putative protein containing 334 amino acid residues. Blast against the current GenBank DNA and protein sequence database did not reveal significant homology with any known proteins. RT-PCR shows that GDR1 mRNA level increased in the differentiated BT-325 cells after being treated with RA. The different expression patterns of GDR1 mRNA in human tissues have been detected through the multiple tissue Northern blot hybridization. 相似文献
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Guangwei Du Yan Zhou Jianhe Chen Junhua Wang Bin Yin Jiangang Yuan Boqin Qiang 《科学通报(英文版)》2000,45(7):620-625
Thembl (muscleblind) gene ofDrosophila encodes a nuclear protein which contains two Cys3His motifs. The mutation ofmbl gene will disturb the differentiation of all theDrosophila’s photoreceptors. Primers have been designed according to human EST086139, which is highly homologous tombl gene. Human fetal brain cDNA library has been screened and a novel cDNA clone has been obtained. The 2595 bp cDNA, designatedMBLL (muscleblind-like), contains an open reading frame which encodes 255 amino acids and has 4 Cys3His motifs (GenBank Acc. AF061261). The amino acids sequence shares high homology toDrosophila’s mbl. The Northern blot and RNA dot blot hybridization of 43 human adult tissues and 7 fetal tissues show thatMBLL is a widely expressed gene, but the expression amounts differ in these tissues. 相似文献
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cDNA fragment of the gene (dehydration induced,di1) of wheat (Triticum aestivum. L) induced by 30% PEG-6000 (−1.13 MPa) treatment was isolated with mRNA differential display technique. Northern blot analysis
showed that the expression ofdi1 gene improved at 10 h reached the highest at 48 h under 30% PEG-6000 treatment. cDNA fragment ofdi1 gene has been cloned and sequenced (211 bp). DNA sequence analysis shows that there is no homologue in GenBank todi1 cDNA. 相似文献
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Min Zhang Long Yu Qiang Tu Peirong Hu Qi Zhang Anding Bi Chunling Jiang Shouyuan Zhao 《科学通报(英文版)》1999,44(9):799-799
An EST (gb/AA115239) with high identity to the mouse cytokine signal transduction inhibitor genemmSOCS-2 was selected in GenBank EST database by the homologous screening method. The cDNA with the same sequence of the EST was got
in human placenta cDNA library by PCR and a 1011 bp cDNA fragment was selected using above cDNA as probes to perform walking
hybridization in placenta cDNA library. The cDNA fragment contains one 594 bp open reading frame (ORF) which encodes 198 amino
acid residues. It was proved to be novel after NCBl database screening. Homology comparison showed that this gene has 93%
identity tommSOCS-2 at the amino acid level and it has high identities to other related genes in SH2 domain and SOCS box, so it was namedhumSOCS-2 and the accession number in GenBank is gb/AF020590. The expression analysis showed that the gene is expressed obviously higher
in prostate than in other 15 human tissues. 相似文献
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取连续培养的骨髓基质细胞并提取其mRNA,合成cDNA后,收集>400bp的片段,连接到真核表达载体pcDNA3.0上,构建人胎儿骨髓基质细胞真核cDNA质粒文库,并以PCR的方法从文库中扩增出编码人IL-6、SCF的基因序列,随机对文库克隆进行测序,得到了3个新基因EST,其中2个在GenBank的登录号分别为AF244998及AF244999。 相似文献
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The fusion gene of actin (cDNA ofChlamydomonas reinhardtii) and green fluorescence protein (gfp) had been constructed into two expression vectors which could be expressed inE. coli and tobacco suspension cells BY2. The correct expression was observed inE. coli and BY2 with a fluorescence microscopy. The fusion protein, which took part in the membrane skeleton, was mainly located
peripherally along the membrane, specially the fusion protein was distributed around nucleus and cell plate, while the fusion
protein also forms F-actin in the cell. The fusion protein was purified from Bl21plus by ammonium sulfate fractionation, ion
exchange chromatography and hydrophobic interaction chromatography. The purified production could polymerize into F-actin
when the actin polymerizing buffer was added. It was demonstrated that the characteristics and function of actin inChlamydomonas was similar with those of animals and higher plants. 相似文献
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ECBP21 is an extracellular calmodulin-binding protein which was first detected and purified from extracellular extracts of
suspension-cultured cells of Angelica dahurica. The purified protein was electroblotted onto PVDF membrane and the amino acid sequences from 1 to 20 were determined. Using
degenerate oligonucleotides of the sequence, a full-length cDNA coding for ECBP21 was isolated by a combination of RT-PCR
and 5′-RACE cloning. The cDNA contains 947 nucleotides and codes for a precursor protein of 216 amino acids. The N-terminal
1–25 amino acid sequence is a predicted signal peptide and the other 26–216 amino acid sequence is a mature peptide. The 26–45
amino acid sequence shows identity with the N-terminal amino acid sequence of purified ECBP21 from Angelica dahurica. The fragment of encoding the mature protein was cloned into pET-28b(+) and transformed into E. coli BL21(DE3). A protein with relative molecular mass 21 ku was expressed in E. coli. Using a biotinylated-CaM gel overlay technique, the expression protein was tested for its ability to bind CaM. The results
indicated that the expression protein is a Ca2+-dependent CaM-binding protein. Thus, these results further defined the cDNA clone for ECBP21. This work laid a foundation
for elucidating biological functions of ECBP21 by using molecular biological means. 相似文献
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Qiang Zhang Lin Chen Junkui Ai Lee Chung Lili Liang Ming Tong Zhiwen Zhang Yanqun Na Yinglu Guo 《科学通报(英文版)》2001,46(19):1622-1626
After the renal cell carcinoma related novel gene fragmentGYLZ-RCC18 was cloned by using suppression subtractive hybridization (SSH), we used the SMART RACE technology to clone the full length
ofGYLZ-RCC18 and performed chromosome location by the FISH method. RT-PCR was used to detect the expression of the first reading frame
ofGYLZ-RCC18 in different stages and grades of renal cell carcinoma tissue and other tissues. Also we transfected the antisense oligonucleotide
ofGYLZ-RCC18 to renal cell carcinoma cell line GRC-1, and analyzed the proliferation activity, growth speed, apoptosis and mortality changes
in GRC-1. The results show that the full length ofGYLZ-RCC18 (GenBank accession No.: BE825133) cDNA is about 3.5 kb long which is located at No. 14 chromosome.GYLZ-RCC18 has a higher expression in higher grades and stages of renal cell carcinoma than in the lower ones. The expression ofGYLZ-RCC18 in renal cell carcinoma was much higher than that in normal kidney and other tissues. After transfection ofGYLZ-RCC18 antisense oligonucleotide, the mortality of GRC-1 increases evidently, the proliferation activity and growth speed were inhibited
remarkably at the same time. Also the antisense oligonucleotide can induce the apoptosis of GRC-1 all through the observation
time. Our results indicated thatGYLZ-RCC18 is an important novel gene related to renal cell carcinoma. Its overexpression would stimulate the growth and proliferation
activity and plays an antidead and antiapoptosis effect in renal cell carcinoma. Transfection of antisense oligonucleotide
could inhibit the generation and development of renal cell carcinoma. The study provides a new clue for the research of renal
cell carcinoma, and also provides an instruction for special genetic diagnosis and the therapy of renal cell carcinoma. 相似文献
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The expression of immediate early gene plays a pivotal role in rat hepatocyte proliferation from G0 to G1 phases and the progression through G1 phase of the cell cycle within several hours after 2/3 hepatectomy. We investigated the different gene expressions within
1 h after 2/3 hepatectomy by representational difference analysis. Sequence analysis indicated thatPC3 induced by NGF was a kind of immediate early gene and might be correlated with liver regeneration. Moreover, we found that
2/3 hepatectomy could induce the expressing ofPC3 mRNA by Northern blot with a peak 1–2 h after surgery. In primary cultures of rat hepatocytes, addition of EGF resulted in
rapid and transient induction ofPC3 mRNA. It was first reported thatPC3 gene belongs to immediate early gene associated with liver regeneration. 相似文献
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Apoptosis of vascular endothelial cells (VEC) has been induced by deprivation of survival factors (aFGF and serum) and by
rattlesnake venom. The expression ofbcl-2 gene has been examined by Northern blotting in the two apoptosis inducing systems. Our results show that the expression ofbcl-2 has not been detected in normal culture cells and in apoptotic cells induced by deprivation of survival factors. But in apoptotic
cells induced by rattlesnake venom (10 ng/mL), the expression ofbcl-2 increases, and its mRNA exhibits two bands. The data first suggest that increasing expression and splitting ofbcl-2 mRNA may play an important role in apoptosis of VEC induced by rattlesnake venom, and this finding is helpful to understanding
the role ofbcl-2 in regulation of apoptosis. 相似文献
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The cDNA of soluble human tumor necrosis factor receptorⅠ(sTNFRI) was inserted into fusion-protein expression plasmid pIGF of A. niger to construct fusion expression vector pHBC containing a KEX2 like protein processing site designed on the fusion position. Extracellular protease-deficient strain of A. niger 3.795-1-23 was transformed with pHBC. Positive clone was estimated by Southern hybridization. SDS-PAGE for protein produced by recombinant strain showed the distinctive expression band. Western blotting indicated that the secreted protein had immunoactivity of sTNFRI. 相似文献
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Junhua Xiao Lanlan Yin Jianmin Li Hu Zu Zuomin Zhou Baige Zhao Jiahao Sha 《科学通报(英文版)》2002,47(11):896-901
Using cDNA microarray hybridization from a human testicular cDNA library, one gene exhibiting ten-fold difference at expression
level between adult and embryo human testes was cloned and named NYD-SP9, which was believed to be involved in spermatogenesis. Southern blot hybridization results showed that NYD-SP9 expressed highly in testis but low in ovary. Protein motif analysis of this cDNA sequence revealed a cluster of phosphorylation
sites, indicating its potential involvement in signal pathways during spermatogenesis. Furthermore, one transmembrane helix
was predicted in N-terminal region, indicating that putative NYD-SP6 may be served as a transmembrane protein. The proximity
of these potential phosphorylation sites to each other indicates that there may be interaction among these sites to regulate
spermatogenesis. These findings suggested that protein kinase NYD-SP9 might play a role in male germ cell differentiation. 相似文献
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The role of PKCα in human breast cancer cell proliferation and expression ofcyclinD1 andCDK4 has been investigated using inhibition ofPKCα expression by its antisense RNA. WhenPKCα expression was inhibited the rate of cell proliferation decreased apparently and the levels ofcyclinD1 andCDK4 mRNA were lower than the control. The results showed thatPKCα, a key member of signal transduction system, played an important role in human breast cancer cell proliferation and had a
close relationship with expression ofcyclinD1 andCDK4 which control start of cell cycle. 相似文献