人p16~(INK4) cDNA探针在非小细胞肺癌研究中的应用

制备总RNA ,RT PCR克隆p16 INK4 cDNA ,测序验证 ,制备探针 .进行PCR产物Southern杂交检测非小细胞肺癌组织标本中p16 INK4 基因第二外显子阴性杂交率为 12 .9% (4 / 31) .原位杂交显示p16 INK4 基因转录阴性率为2 2 .6 % (7/ 31) .结果说明克隆的p16 INK4 cDNA是正确的 ,可用于临床基因诊断 ,p16 INK4 基因变异及表达在非小细胞肺癌的发生、发展中起作用     

Identification of differentially expressed genes in esophageal cancer through SSH in combination with high throughput reverse Northern screening

To understand the molecular mechanisms of carcinogenesis of esophagus and to isolate genes with different expression levels in esophageal cancer, suppression subtractive hybridization (SSH) was combined with PCR-based cDNA synthesis and reverse Northern on the cancer tissues and matched almost normal mucosa using 5 microgram of total RNA as starting marterial. Eight genes were found expressed differentially in esophageal cancer, in which 5 were known genes and 3 were novel ones; and 6 were down-regulated in cancer tissues, while 2 were up-regulated; 6 were of mid-high abundance and 2 were of low abundance in esophagus. The results revealed that alteration in expression level of multiple genes underlied the initiation and development of esophageal cancer. The differentially expressed genes identified in this study such as liporcotinⅠ, cystatin A, cystatin B, cytokeratin 13 may play roles in dedifferentiation, transformation and malignant proliferation of esophageal cancer. The combination of SSH with PCR-based double- strand cDNA synthesis and high throughput reverse Northern screening is an efficient way to isolate differentially expressed genes from microgram of total RNA.     

p53基因在小鼠胚胎发育过程中的表达

以E9日龄至E14日龄昆明种正常小鼠胚胎为材料,利用质粒扩增的、地高辛标记的基因探针在组织切片上进行DNA-mRNA 分子原位杂交,研究了p53基因在小鼠胚胎发育过程中的表达.结果表明, p53基因不参与E9和E10日胚胎发育中的器官原基形成,参与器官的进一步分化成熟过程.这些器官主要有眼、脑、心、肺、脊柱和面颌骨,肝组织的发育与其无关; p53基因一方面参与胚胎发育中的细胞周期调控,另一方面也参与了某些与细胞周期无关的过程;不同的器官有不同的细胞周期调控机制.     

酶学信号扩增方法定量检测生物素探针杂交

将靶核酸固定于微孔滴定板上，用化学方法标记的生物素探针进行杂交，采用底物－辅助因子循环信号扩增系统对杂交结果进行检测，结果表明用该信号检测系统的检验灵敏度比常用显色底物的检测灵敏度高１０倍。     

转基因马铃薯植株中外源基因PCR扩增和拷贝数测定

本文报道一种简便、快速、可靠，同时又适合大量转基因植株中外源基因测定的ＰＣＲ检测技术，同时改进了转基因植物中总ＤＮＡ提取方法，并且用窄缝转移杂交测定了转基因植株中外源基因拷贝数。     

柴胡与高寒藏药-川西獐芽菜科间体细胞杂交

川西獐芽菜原生质体用强度为 2 6 0 μw/cm2 紫外线照射 1、2、3min后 ,与狭叶柴胡原生质体在PEG诱导下融合 .对融合再生的 4 6个单细胞克隆进行杂种形态学、染色体、同功酶分析 ,结果表明 ,它们中有 32个为体细胞杂种细胞系 .11个克隆在 3个月时再生出完整小植株 .     

后殖民理论视角下赛译《水浒传》的杂合特征研究

后殖民翻译理论关注隐藏在译文背后的两种文化间的权力争斗和权力运作,认为翻译既是殖民化的途径,也是解殖民化的工具。翻译中的杂合是语言文化交流的必然产物,有利于彰显文化差异、消解文化霸权。赛译水浒对异质性话语的保留使译本呈现出明显的杂合特征。本文拟在后殖民理论视角下,分别从字词、句法、篇章和文化四个层面对赛译水浒的杂合特征进行系统的阐释,旨在发掘赛译水浒的文化价值和文学价值,并阐明其对当下中国文学对外传播过程中的启迪意义。     

Chymotrypsin gene expression during the intermolt cycle in the shrimpPenaeus vannamei (Crustacea; Decapoda)

InPenaeus vannamei, chymotrypsin is present as two isoenzymes in the hepatopancreas. The enzyme has been localized in F-cells by immunocytochemistry using a specific antibody. By in situ hybridization, with a 510 pb cDNA probe encoding for the first 170 amino acids of the shrimp chymotrypsin, mRNA was localized in the same cells. Gene expression was followed during the intermolt cycle by measuring changes in specific activity in crude extracts, and by the estimation of mRNA levels by Northern blots using the same probe. The increase in specific activity in premolt is preceded in early premolt by an increase in the amount of chymotrypsin mRNA. A second increase is observed in postmolt, suggesting a different mode of regulation of gene expression.