成熟大豆种子中 mRNA 的菌落杂交研究。

A colony hybridization study of mRNA in maturating soybean seeds.

机构信息

Institute of Genetics and Selection of Industrial Microorganisms, Dorozhnaya 8, 113545, Moscow, USSR.

出版信息

Plant Cell Rep. 1982 Dec;1(6):267-9. doi: 10.1007/BF00272636.

Abstract

Polysomal poly(A)RNA was transcribed by avian myeloblastosis virus (AMV) reverse transcriptase and double-stranded complementary DNA (cDNA) was then synthesized and cloned in Escherichia coli C600 by (dG)-(dC) tailing in the PstI site of pBR322. A bank of mRNA sequences from maturating soybean seeds (average weight 200 mg) was obtained. (32)P-cDNA based on mRNA from various organs of the plant (leaves, axes, seeds) was synthesized. The clones were hybridized with different kinds of (32)P-cDNA on nitrocellulose filters. Approximately 13% of the clones were specific for the maturating stage, and more than a half the clones gave a positive result with all kinds of cDNA.

摘要

多核糖体多聚（A）RNA 由禽成髓细胞瘤病毒 (AMV) 逆转录酶转录，双链互补 DNA (cDNA) 随后通过 pBR322 的 PstI 位点的 (dG)-(dC) 加尾在大肠杆菌 C600 中合成并克隆。从成熟大豆种子（平均重量 200 毫克）获得了一批 mRNA 序列库。（32）P-cDNA 基于来自植物不同器官（叶片、轴、种子）的 mRNA 合成。将克隆与不同类型的（32）P-cDNA 在硝酸纤维素滤膜上杂交。大约 13%的克隆对成熟阶段具有特异性，超过一半的克隆与所有类型的 cDNA 都有阳性结果。