大豆不同组织mRNA体外翻译产物2D PAGE的比较

2D PAGE COMPARISON OF IN VITRO TRANSLATION PRODUCTS OF mRNAs FROM DIFFERENT TISSUES OF SOYBEAN

从大豆的叶片、种子、茎阳根制备了未降解的、具有生物活性的Poly(A)^+mRNA。建立了高效翻译的麦胚无细胞系统。通过对不同组织mRNA翻译产物双向聚丙烯酰胺凝胶电泳图谱(2D图谱)的比较,找出了每种组织的专一性或高丰度的蛋白质斑点。这些结果有助于克隆组织专一性的DNA调控序列和转基因植物的研究。

Undegrated Poly(A)^+mRNA with biological activity were isolated from the leaves, seeds, stems and roots of soybean, A high efficient translation system was constructed from wheat germ, The differences between four mRNA populations were revealed by analysis of their wheat germ translation products on two-dimensional isofocusing/SDS gel electrophcresis, and some of tissue-specific and abundant protein spots for each of tissueerew found out by comparison of their 2D patterns.The 2D pattern of seed mRNA translation products was markedly different from that of other three tissues, Seven seed-specific protein spots were found in the 58—66/6.0—7.0 region. Many similarities were present in the different translation products of leaf, stem, and root mRNAs. In leaf, spot 26/8.4 was the leaf-specific and most abundant protein, a group of four leaf-speficic proteins were in 52/6.0—6.8 region, and the spot 76/4.8 was much denser and larger than corresponding spots in other tissue 2D patterns. These results have a implication in Searching tissue-specific regulatory DNA sequences and engineering tissue-specifically expressed transgenic plants.