Detection of a gene which is forming a tree stem

目前，对形成树干的基因的研究还没有现成的方案。除了在其他生物材料的基因工程中开发的常规方法之外，这些研究还需要新的方法。以下两个是为研究树干中的基因而开发的。首先，用于提取树茎中的基因（如RNAS）的方法。第二，检测形成树干的基因的方法。本研究对这几点进行了研究。首先，通过几种不同的提取方法获得了RNA组分。如果分离到完整的RNA，则主要通过紫外光谱和琼脂糖凝胶电泳来检查RNA质量。在去除蛋白质和/或果胶杂质后，它得到改善。经poly（A+）RNA分级分离后，合成的cDNA最大链长约为2.2kb，包含81.5Kdalton多肽。其次是外源凝集素基因。树中的凝集素基因由一个基因家族组成，并且表现出器官特异性表达。这些器官特异性基因家族可能参与了该物种在进化过程中由树栖向草栖的转变。因此，根据氨基酸序列，用合成的混合探针对国槐凝集素基因进行了检测。该基因似乎在种子RNA中检测到，而在茎制备物中没有观察到基因。在新开发的两种方法中，第一种方法已基本确定，但第二种方法，即基因检测，还需要进一步研究。

At present, no protocol is available in the studies on genes which are forming tree stems. New methods are needed for those studies in addition to the conventional methods which have been developed in genetic engineering in other biological materials. Following two should be originally developed for the studies on the genes in a tree stem. First, the method for the extraction of the genes, as RNAS, in a tree stem. Secondary, the method for detecting genes which are forming tree stems. These points are examined in this study.Concerning to the first place, RNA fractions were obtained by several extraction methods. The RNA quality was examined by mainly using UV spectra and agarose gel electrophoresis, if intact RNAs were isolated. It was improved after proteinous and/or pectic impurities had removed. The cDNAs synthesized from such RNAs after poly(A+)RNA fractionation, showed maximal chain length about 2.2kb, which covered 81.5Kdalton polypeptides. Concerning to the second place, it was on a lectin gene. Lectin genes in a tree consist of a gene family, and show organ specific expression. Such organ specific gene family had probably been involved in the conversion of arboreal to herbaceous habits during the evolution. Thus, lectin genes in Sophora japonica(Sophora contains habits, tree, shrub and herb) were examined by using a synthetic mixed- probe according to the amino acid sequences. The gene seemed to be detected in the seed RNAs, while no gene was observed in the stem preparation. In the two methods to be newly developed, the first one was almost confirmed, but further studies are needed for the second one, i.e. gene detection.