植物基因表达的组织原位杂交和原位组织化学分析技术的改进  

IMPROVED APPROACHES OF IN SITU HYBRIDIZATION AND IN SITU ENZYME HISTOCHEMISTRY FOR  GENE EXPRESSION ANALYSES IN PLANTS

作　　者：刘庆法[1,2,3] 郝峥嵘[1,2,3] 沈大棱[1,2,3] 

机构地区：[1]复旦大学遗传研究所 [2]西北农业大学农学系 [3]新疆农业科学院核技术生物技术研究所

出　　处：《Acta Botanica Sinica》1998年第6期580-584,共5页Acta Botanica Sinica（植物学报：英文版）

摘　　要：改良了组织原位杂交和原位酶组织化学分析的方法。主要改进有：原位杂交采用简化的ＦＡＡ固定程序，常规石蜡包埋，切片时采取液氮深冷冻；以随机引物法标记的ＤＮＡ代替转录标记的ＲＮＡ作探针，在保湿盒中进行杂交，而不用矿物油覆盖。组织化学分析中用含铁氰化钾的显色液进行ＧＵＳ染色后再包埋、切片的程序，代替先包埋、切片再显色的常规方法，可最大限度地保持酶活性的真实性。Improved methods for both in situ hybridization and in situ enzyme histochemistry were described. The procedures for both methods have been significantly simplified by omitting some unnecessary treatments and substituting the cumbersome and laborious techniques, and the reliability of in situ histochemistry was increased by reversing the operations of Block and Debrouwer's procedure. The improved steps are: Instead of the conventional fixation, a simplified FAA procedure by adding liquid nitrogen onto the embedded tissues during sectioning to ensure high quality of the sections;labeled DNA by randompriming, other than labeled RNA by transcription, was used as probes in hybridization, which was conducted in a moisturesaturated plastic chamber other than emerging in mineral oil. The improved procedure for tissue in situ histochemical study was that the GUS coloration was carried out before fixation, embedding and sectioning, which was different from the procedure as described by Block and Debrouwer.

关 键 词：原位杂交 原位组织化学 基因表达 植物 

分 类 号：Q94－33[生物学—植物学]