从原核生物海洋原绿球藻（原绿藻门）基因组中扩增的随机扩增多态性DNA片段的测序。

Sequencing of RAPD fragments amplified from the genome of the prokaryote Prochlorococcus marinus (prochlorophyta).

作者信息

Lorenz M, Partensky F, Börner T, Hess W R

机构信息

Department of Biology, Humboldt University Berlin, Germany.

出版信息

Biochem Mol Biol Int. 1995 Jul;36(4):705-13.

PMID:8528133

Abstract

DNA fingerprint patterns from the chlorophyll a and b containing prokaryote Prochlorococcus marinus were generated with the RAPD technique using two primers derived from repetitive sequence motifs, [(GATA)4 and M13] and a random primer (OPB-10]. Five RAPD fragments were reamplified, cloned and sequenced. The clones M13/1300 and OPB-10/1100 contained open reading frames, whereas the (GATA)4 fragments were interrupted by stop codons in all frames indicating their noncoding function and possessed a high AT score of 63% and 71%, respectively. With the two (GATA)4 clones and the M13/300 clone strain-specific signals were obtained in a Southern blot analysis of various Prochlorococcus strains. Clones M13/1300 and OPB-10/1100, containing the ORFs, produced RFLPs between the strains analyzed. All RAPD fragments are represented as single copy in the genome of Prochlorococcus.

摘要

利用随机扩增多态性DNA（RAPD）技术，使用源自重复序列基序的两种引物[（GATA）4和M13]以及一种随机引物（OPB - 10），生成了含有叶绿素a和b的原核生物海洋原绿球藻的DNA指纹图谱。对五个RAPD片段进行了再扩增、克隆和测序。克隆M13/1300和OPB - 10/1100含有开放阅读框，而（GATA）4片段在所有阅读框中均被终止密码子打断，表明它们具有非编码功能，并且其AT含量分别高达63%和71%。利用两个（GATA）4克隆和M13/300克隆，在对各种海洋原绿球藻菌株的Southern印迹分析中获得了菌株特异性信号。含有开放阅读框的克隆M13/1300和OPB - 10/1100在分析的菌株之间产生了限制性片段长度多态性（RFLP）。所有RAPD片段在海洋原绿球藻基因组中均以单拷贝形式存在。