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RAPID DETECTION OF ONCOGENE AMPLIFICATION IN PARAFFIN SECTIONS OF BREAST CARCINOMAS USING QUANTITATIVE DIFFERENTIAL PCR AND FLUORESCENT DNA TECHNOLOGY
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作者 An Hanxing Niederacher D Beckmann MW 《Chinese Medical Journal》 SCIE CAS CSCD 1995年第3期68-69,共2页
Gene amplification is a common mechanism of oncogene activation and contributes to tumor progression. Analysis of such genetic alterations are relevant to the understanding of tumor genetics and could provide prognost... Gene amplification is a common mechanism of oncogene activation and contributes to tumor progression. Analysis of such genetic alterations are relevant to the understanding of tumor genetics and could provide prognostic information for the individual patient. Standard analytical approaches using Southern blot and slot blot require a large amount of good 展开更多
关键词 pcr GENE DNA RAPID DETECTION OF ONCOGENE AMPLIFICATION IN PARAFFIN SECTIONS OF BREAST CARCINOMAS USING QUANTITATIVE differential pcr AND FLUORESCENT DNA TECHNOLOGY
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Isolating cDNA clones from rice induced by Magnaporthe grisea using PCR based differential screening method
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作者 DONG Haitao, DONG Jixin, WU Yuliang, HE Zuhua,and Li Debao,Biotechnology Institute,ZheJiang Agri Univ, Hangzhou 310029, China 《Chinese Rice Research Newsletter》 1998年第3期1-2,共2页
The differential hybridization technique hasbeen widely used to identify genes that are dif-ferentially expressed.However,this approachhas several drawbacks.First,the screeningprocedures are rather labor-intensive and... The differential hybridization technique hasbeen widely used to identify genes that are dif-ferentially expressed.However,this approachhas several drawbacks.First,the screeningprocedures are rather labor-intensive and time-consuming.Second,the amount of phageDNAs transferred onto the two filters may notbe equivalent,which leads to an inaccurate se-lection of a positive clone.Third,isolation ofphage DNA is slow and cumbersome.Here,aPCR based differential screening method that 展开更多
关键词 cDNA pcr Isolating cDNA clones from rice induced by Magnaporthe grisea using pcr based differential screening method
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Primarily screening and analyzing ESTs differentially expressed in rats' primary liver cancer 被引量:9
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作者 Dandan Liu Lijuan Zhi +9 位作者 Mingxia Ma Dan Qiao Meijuan Wang Yawei Wang Baijie Jin Anqi Li Guting Liu Yiqing Zhang Yanyan Song Hongxu Zhang 《Chinese Journal of Cancer Research》 SCIE CAS CSCD 2013年第1期71-78,共8页
Objective: To screen and analyze key express sequence tags (ESTs) which were differentially displayed in every period of SD rats' primary hepatic carcinoma and reveal the molecular mechanism of carcinogenesis. Met... Objective: To screen and analyze key express sequence tags (ESTs) which were differentially displayed in every period of SD rats' primary hepatic carcinoma and reveal the molecular mechanism of carcinogenesis. Methods: Using diethylnitrosamine (DENA) as a cancerigenic agent, animal models with different phases of primary hepatic cancer were constructed in SD rats. Rats were respectively sacrificed at d 14, d 28, d 56, d 77, d 105 and d 112 after the rats received DENA by gavage, then the livers were harvested. One part of the livers was classified according to their pathological changes, while the other was reserved for molecular mechanism studies on hepatocarcinogenesis. The differentially expressed genes were isolated from both normal and morbid tissues by mRNA differential display technique (DDRT-PCR). After the fragments were sequenced, bioinformatics were .used to analyze the results. Results: Twelve differentially expressed cDNA fragments were obtained. Nine fragments had the homology with known cDNA clones, especially EST-7 was similar to BN/SsNHsdMCW mitochondrion gene and the identity was 100% which suggested EST-7 may be the part of BN/SsNHsdMCW mitochondrion gene. In contrast, other three fragments (EST-1, EST-3 and EST-5) had extremely low identity to any genes registered in GENBANK databases. Conclusions: BN/SsNHsdMCW mitochondrion gene was expressed in different periods of hepatocarcinogenesis. Moreover, EST-I, EST-3 and EST-5 were suggested to contribute to the development of rat hepatocarcinogenesis, and thus may be candidates of new targets of oncogenes or cancer suppressor genes. 展开更多
关键词 Animal models of primary liver cancer DDRT-pcr differential display reverse transcription pcr ESTs (express sequence tags) mitochondrion gene
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Differential Gene and Protein Expression in Soybean at Early Stages of Incompatible Interaction with Phytophthora sojae 被引量:1
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作者 LI Yong-gang YANG Ming-xiu +3 位作者 LI Yan LIU Wen-wen WEN Jing-zhi LI Yong-hao 《Agricultural Sciences in China》 CAS CSCD 2011年第6期902-910,共9页
Soybean root and stem rot caused by Phytophthora sojae is a destructive disease worldwide. Using genetic resistance is an important and major component in the integrated pest management of this disease. To understand ... Soybean root and stem rot caused by Phytophthora sojae is a destructive disease worldwide. Using genetic resistance is an important and major component in the integrated pest management of this disease. To understand molecular mechanisms of root and stem rot resistance in soybeans, the gene and protein expression in hypocotyls and stems of variety Suinong 10 carrying resistance genes Rps1a and Rps2 was investigated by using mRNA differential display reverse transcription PCR and two-dimensional electrophoresis at 0, 0.5, 1, 2, and 4 h after inoculation with P. sojae race 1. The results of the comparison of gene and protein expression showed that at least eight differential fragments at the transcriptional level were related to metabolic pathway, phytoalexin, and signal transduction in defense responses. Sequence analyses indicated that these fragments represented cinnamic acid 4-hydroxylase gene, ATP b gene coding ATP synthase b subunit and ubiquitin-conjugating enzyme gene which upregulated at 0.5 h post inoculation, blue copper protein gene and UDP-N-acetyl-a-D-galactosamine gene which upregulated at 2 h post inoculation, TGA-type basic leucine zipper protein TGA1.1 gene, cyclophilin gene, and 14-3-3 protein gene which upregulated at 4 h post inoculation. Three resistance-related proteins, a-subunit and b-subunit of ATP synthase, and cytochrome P450-like protein, were upregulated at 2 h post inoculation. The results suggested that resistance-related multiple proteins and genes were expressed in the recognition between soybean and P. sojae during zoospore germination, penetration and mycelium growth of P. sojae in soybean. 展开更多
关键词 Phytophthora sojae resistance mechanism incompatible interaction mRNA differential display reverse transcription pcr two-dimensional electrophoresis
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mRNA differential display on gene expression in settlement metamorphosis process of Ruditapes philippinarum larvae
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作者 卢素敏 Bao Zhenmin +3 位作者 Hu Jingjie Hu Xiaoli Mu Chunhua Fang Jianguang 《High Technology Letters》 EI CAS 2008年第3期332-336,共5页
The mRNA differential display (DDRT-PCR) technique was adopted to find out the genes related tosettlement metamorphosis development process of Ruditapes philippinarum larvae.In this study,we haveobtained three hundred... The mRNA differential display (DDRT-PCR) technique was adopted to find out the genes related tosettlement metamorphosis development process of Ruditapes philippinarum larvae.In this study,we haveobtained three hundred and forty-six amplification bands in total from pediveliger larvae,veliger larvae,eye spot larvae and post-larvae.Sixty-five out of three hundred and forty-six bands are distinctly differen-tial display from band pattern,which can be put into four groups,standing for different expression char-acters.Sixteen differential display bands were cloned,sequenced and analyzed and nine different se-quences are obtained in the study.Three sequences have higher similarity to the cDNAs deposited indatabase and three are very similar to the rDNA of other species,considered as the rDNA of Ruditapesphilippinarum.The rest three sequences are found to be novel sequences after analyzed.Their accessionnumbers are AY916799,AY916798,and AY916797 respectively.We thought the novel sequences arepossibly relevant to the early embryo development of Ruditapes philippinarum larvae and can provide somefundamental understandings that are helpful for the improvement of scallop seed raising industry. 展开更多
关键词 DDRT-pcr (mRNA differential display pcr differential gene expression Larvae development settlement metamorphosis Ruditapes philippinarum
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Inducible Expression of Translation Elongation Factor 1A Gene in Rice Seedlings in Response to Environmental Stresses 被引量:13
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作者 李子银 陈受宜 《Acta Botanica Sinica》 CSCD 1999年第8期800-806,共7页
Differences of gene expression between salinity_stressed and control rice ( Oryza sativa L. ssp. indica ) cultivar “Zhaiyeqing 8' were compared using differential display PCR (DD_PCR) technique. Sequence an... Differences of gene expression between salinity_stressed and control rice ( Oryza sativa L. ssp. indica ) cultivar “Zhaiyeqing 8' were compared using differential display PCR (DD_PCR) technique. Sequence analysis of one salt_inducible cDNA clone revealed that this clone represented a new member of rice translation elongation factor 1A (eEF1A) gene family and was tentatively named REF1A. Northern blot hybridization using REF1A fragment as a probe was performed to investigate the expression of rice translation elongation factor 1A gene in response to various environmental factors. It was observed that expression of the eEF1A gene in rice shoots was dramatically induced by salinity stress or exogenous application of abscisic acid (ABA). The induction of this gene by ABA stress occurred more quickly than that by salinity stress. In addition, expression of rice translation elongation factor 1A gene was also induced by drought (15% PEG6000), cold (4 ℃) or heat_shock (37 ℃) stresses. The results suggested that the induction of translation elongation factor 1A gene expression by environmental stresses might reflect the general adaptive response of rice plants to the adverse circumstances. 展开更多
关键词 RICE differential display pcr Translation elongation factor 1A Environmental factors differential expression
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Effect of Epinephrine on the Settlement and Metamorphosis of Manila Clam Larvae 被引量:2
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作者 LU Sumin BAO Zhenmin +1 位作者 LIU Hui FANG Jianguang 《Journal of Ocean University of China》 SCIE CAS 2006年第2期141-145,共5页
Chemical inducement and DDRT-PCR (differential display reverse transcription PCR) are adopted to investigate the effect of epinephrine (EPI) on the settlement and metamorphosis of Manila clam larvae. Chemical indu... Chemical inducement and DDRT-PCR (differential display reverse transcription PCR) are adopted to investigate the effect of epinephrine (EPI) on the settlement and metamorphosis of Manila clam larvae. Chemical inducement shows that EPI has an effect to some extent on the metamorphosis of Manila clam larvae at all concentrations and in all treatments designed. The most significant result of inducement is obtained at the concentration of 10^-6 tool L^-1 and for 4 h. DDRT-PCR using six primer pairs shows that the gene expression pattern is quite different between EPI treatment and the control. Three hundred and forty-three amplification bands are obtained in total, among which, 67 (19.53%) are differentially appeared. Therefore, EPI has an effect on the gene expression of the eye spot larval Manila clam. It can be hypothesized that EPI is a settlement and metamorphosis inducer for Manila clam. EPI may lead to larvae settlement and metamorphosis by binding to the receptors on the membrane and then changing the gene expression of larvae cells. 展开更多
关键词 epinephrine (EPI) differential display reverse transcription pcr (DDRT-pcr settlement and metamorphosis Manila clam Ruditapes philippinarum
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Differential Display of Cotton cDNAs Expressed by Salicylic Acid Induction 被引量:1
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作者 李骥 赵广荣 刘进元 《Tsinghua Science and Technology》 SCIE EI CAS 2003年第4期498-501,共4页
Salicylic acid (SA) is very important in systemic acquired resistance and hypersensitive response in plant defense, and yet its role is not fully understood. This study seeks to clarify the mechanism of SA induced res... Salicylic acid (SA) is very important in systemic acquired resistance and hypersensitive response in plant defense, and yet its role is not fully understood. This study seeks to clarify the mechanism of SA induced resistance in cotton. Total RNA was extracted from low gossypol cultivated cotton seedlings treated with exogenous SA and subjected to fluorescent differential display PCR (FDD PCR). Seven cDNA fragments were selected from the total ten differential bands. Comparison with Genbank database shows that all seven cDNA sequences are newly discovered in cotton. However, they share high amino acid identity to some registered cDNAs. Among them, three of the cDNAs could be predicted to encode basic chitinase, penicillin binding 6 b precursor and ATP dependent DNA helicase RecG, while the functions of the other four cDNAs are undetermined. Dot blot analysis demonstrates that the expression of five cDNAs in cotton seedlings is induced by SA, while SA induction has a negative effect on the transcript accumulation of the other two cDNAs (E13 and E14). Since SA was previously shown to enhance the resistance to cotton wilt disease, the finding of a basic chitinase gene in cotton expressed by SA induction will provide a new insight into induced disease resistance in cotton. 展开更多
关键词 cotton cDNA salicylic acid induced expression fluorescent differential display pcr (FDD pcr)
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