Synthetic deoxyoligonucleotides as general probes for chloroplast transfer RNA genes.
Synthetic deoxyoligonucleotides as general probes for chloroplast transfer RNA genes.
复制标题
合成脱氧寡核苷酸作为叶绿体转移 RNA 基因的通用探针。
DOI：
10.1093/nar/10.24.8191
复制
发表时间：
1982
影响因子：
14.9
通讯作者：
Hallick,RB
中科院分区：
文献类型：
--
作者：
Nickoloff,JA;Hallick,RB
化学合成脱氧寡核苷酸作为杂交探针检测tRNA基因的效用已被检验。本研究选用叶绿体tRNA基因。利用化学方法合成了高等植物和细叶丑茅叶绿体tRNA基因高度保守区域的脱氧寡核苷酸。这些合成探针已被用于叶绿体dna限制性片段的南杂交检测tRNA基因。这种tRNA基因定位的新方法及其合成的寡核苷酸可能在许多叶绿体基因组中具有普遍的应用价值。这可以通过对凤仙花和菠菜中已知和未知tRNA基因的检测，以及玉米和黄瓜叶绿体dna中未知tRNA基因的检测来说明。已经确定了细叶菊叶绿体tRNAPhegene的精确位置和极性。我们还描述了有关杂交时间、洗涤强度和碱基对错配对杂交信号影响的实验。
The utility of chemically synthesized deoxyoligonucleotides as hybridization probes for the detection of tRNA genes has been examined. Chloroplast tRNA genes were chosen for this study. Deoxyoligonucleotides complementary to highly conserved regions of chloroplast tRNA genes of both higher plants andEuglena graciliswere chemically synthesized. These synthetic probes have been used to detect tRNA genes by Southern hybridizations to restriction fragments of chloroplast DNAs. This new method of tRNA gene mapping and the oligonucleotides synthesized may be of general application to many chloroplast genomes. This is illustrated by the detection of known and unknown tRNA genes ofEuglena gracilisand spinach, and unknown tRNA genes of maize and cucumber chloroplast DNAs. The precise locus and polarity of theEuglena gracilischloroplast tRNAPhegene has been determined. We a1so describe experiments which relate to the effects of the time of hybridization, the stringency of washing, and of base pair mismatches on the hybridization signal.