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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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绵羊单卵反转录平台的优化及应用
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作者 张艳普 孙树春 +3 位作者 田树军 胡媛媛 温兵强 祁昕 《黑龙江畜牧兽医》 CAS 北大核心 2012年第6期3-6,共4页
为了研究发育生物学中单个卵母细胞基因的表达,优化由单卵得到质量良好cDNA的操作步骤,试验应用试剂盒并增加质控环节,得到优质cDNA,同时将cDNA试用于差异显示PCR(DDRT-PCR)。结果表明:该试验方法得到的cDNA稳定性、重复性、完整性都很... 为了研究发育生物学中单个卵母细胞基因的表达,优化由单卵得到质量良好cDNA的操作步骤,试验应用试剂盒并增加质控环节,得到优质cDNA,同时将cDNA试用于差异显示PCR(DDRT-PCR)。结果表明:该试验方法得到的cDNA稳定性、重复性、完整性都很可靠。 展开更多
关键词 单卵 反转录 CDNA 差异显示pcr(ddrtpcr)
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利用DDRT-PCR分析茶树冬季冷驯化过程中基因表达的差异 被引量:7
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作者 梅菊芬 汤茶琴 +1 位作者 徐德良 王新超 《热带作物学报》 CSCD 2011年第4期648-652,共5页
采用DDRT-PCR方法对茶树冬季冷驯化过程中3个样品基因差异表达进行初步研究。结果表明,抗寒性不同的茶树存在基因差异表达。从134条差异片段得到稳定扩增的差异片段31条。根据差异片段的序列设计引物对部分片段进行RT-PCR鉴定假阳性,得... 采用DDRT-PCR方法对茶树冬季冷驯化过程中3个样品基因差异表达进行初步研究。结果表明,抗寒性不同的茶树存在基因差异表达。从134条差异片段得到稳定扩增的差异片段31条。根据差异片段的序列设计引物对部分片段进行RT-PCR鉴定假阳性,得到10个阳性片段,其中6个表达量增加,4个表达量降低。经过BLAST比对分析,这些基因与多种植物基因同源性较高,如nectainⅢ蛋白基因(3)、甘油磷酸二酯磷酸二酯酶蛋白家族基因(48)、微管蛋白特异性C伴侣基因(52)、光合系统Ⅱ蛋白D1基因(80、138)、卵磷脂胆固醇酰基转移酶(147),15、82、134和葡萄的蛋白同源性较高。72经比对没有同源性序列。 展开更多
关键词 茶树 冷驯化 基因表达差异 ddrt-pcr
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