用于检测鸡毒支原体的种特异性生物素化DNA探针。

Species-specific biotinylated DNA probe for the detection of Mycoplasma gallisepticum.

作者信息

Geary S J, Intres R, Gabridge M G

机构信息

Bionique Laboratories, Inc., Saranac Lake, NY 12983.

出版信息

Mol Cell Probes. 1988 Sep;2(3):237-44. doi: 10.1016/0890-8508(88)90007-2.

DOI:10.1016/0890-8508(88)90007-2

PMID:3221885

Abstract

A 5.5 kilobase DNA fragment from an Eco RI digest of the Mycoplasma gallisepticum genome was specific for the detection of M. gallisepticum. This 5.5 kb fragment was initially cloned into bacteriophage lambda gt11 followed by subcloning into the plasmid vector pGEM-3Z. The incorporation of a biotin label was accomplished by utilizing biotin-11-dUTP in a nick translation reaction. This probe, designated pMg6, reacts specifically with M. gallisepticum when tested against various mycoplasma DNAs in Southern blot hybridization analysis. Spot-blot hybridization data indicate the pMg6 is capable of detecting 800 pg of M. gallisepticum DNA.

摘要

来自鸡败血支原体基因组Eco RI酶切片段的一个5.5千碱基DNA片段对鸡败血支原体的检测具有特异性。这个5.5 kb片段最初被克隆到噬菌体λgt11中，随后亚克隆到质粒载体pGEM-3Z中。通过在切口平移反应中使用生物素-11-dUTP来实现生物素标记的掺入。这个名为pMg6的探针在Southern印迹杂交分析中与各种支原体DNA进行检测时，能与鸡败血支原体特异性反应。斑点印迹杂交数据表明pMg6能够检测出800 pg的鸡败血支原体DNA。