TRANSCRIPTION OF BACILLIS-SUBTILIS PLASMID PBD64 AND EXPRESSION OF BACTERIOPHAGE SPO1 GENES CLONED THEREIN
TRANSCRIPTION OF BACILLIS-SUBTILIS PLASMID PBD64 AND EXPRESSION OF BACTERIOPHAGE SPO1 GENES CLONED THEREIN
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DOI：
10.1016/0042-6822(85)90425-8
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发表时间：
1985-01-01
期刊：
影响因子：
3.7
通讯作者：
STEWART, CR
中科院分区：
文献类型：
--
作者：
CURRAN, JF;STEWART, CR
质粒pBD 64，用于在B中克隆的载体。枯草芽孢杆菌具有至少3个实质性转录单位。其中两个包括单个EcoRI、XbaI和BamHI位点，而另一个包括单个BglII位点。相对于pBD 64限制性图谱，这些转录物中的每一种都以逆时针方向合成。在相反方向未检测到转录本。感染的噬菌体SPO1引起这些成绩单大幅下降。在SPO1感染过程中没有检测到新的pBD64转录本。在这些pBD 64位点中的几个位点克隆的各种SPO1基因通过观察其补充SPO1突变体的能力来测试表达。无论片段插入的方向如何，几个中期和晚期基因基本上都表达。由于从载体的转录只能导致1个方向的表达，这表明必需的转录起源于SPO1启动子，因此，SPO1中期和晚期启动子可以在含胸腺嘧啶的DNA中具有活性。
Plasmid pBD64, a vector which is useful for cloning in B. subtilis has at least 3 substantial transcription units. Two of these include the single EcoRI, XbaI and BamHI sites, while the other includes the single BglII site. Each of these transcripts was synthesized in the counterclockwise direction, relative to the pBD64 restriction map. No transcripts were detected in the opposite direction. Infection by bacteriophage SPO1 caused a substantial decrease in each of these transcripts. No new pBD64 transcripts were detected during SPO1 infection. Various SPO1 genes, cloned at several of these pBD64 sites, were tested for expression by observing their capacity to complement SPO1 mutants. Several middle and late genes were expressed substantially, regardless of the orientation in which the fragments were inserted. Since transcription from the vector could cause expression only in 1 orientation, this argues that the necessary transcription originated at SPO1 promoters, and, thus, that SPO1 middle and late promoters can be active in thymine-containing DNA.