概括
DNA旋转酶对于Tn5转移至关重要,需要超绕的DNA. 抑制DNA回旋酶或减少DNA超卷变显著损害了大肠杆菌中的转换效率.
科学领域:
- 分子生物学分子生物学
- 遗传学 是一个遗传学.
- 微生物学 微生物学
背景情况:
- 转换是基因重组的一个关键机制.
- 在各种DNA代谢过程中,DNA超级卷是至关重要的.
研究的目的:
- 阐明DNA旋转酶在Tn5转换中的作用.
- 为了确定DNA超级卷和Tn5转换之间的关系.
主要方法:
- 使用了库默米,一种DNA旋转酶抑制剂.
- 采用了在旋转酶子单元 (gyrA, gyrB) 中发生突变的菌株.
- 引入了一种拓酶I删除来改变DNA超螺旋性.
主要成果:
- DNA 陀螺酶抑制或突变显著降低了 Tn5 转换.
- 减少DNA超级卷,由陀螺酶缺陷引起,转移受损.
- 增加的DNA超卷增强了转换频率,独立于陀螺酶活动.
结论:
- DNA 陀螺酶对于Tn5转换是不可或缺的.
- 超螺旋DNA是Tn5转移的关键基质.
- 这项研究提出了一个有效地将基因转移到大肠杆菌染色体中的系统.
更多相关视频
08:19Generating Transposon Insertion Libraries in Gram-Negative Bacteria for High-Throughput Sequencing
Published on: July 7, 2020
11.6K
11:36Creation of a Dense Transposon Insertion Library Using Bacterial Conjugation in Enterobacterial Strains Such As Escherichia Coli or Shigella flexneri
Published on: September 23, 2017
16.9K
相关概念视频
Transduction
2.6K
Among the three main modes of HGT—transformation, conjugation, and transduction—transduction is unique in that it is mediated by bacteriophages, or bacterial viruses.Transduction occurs in two ways. Generalized transduction occurs during the lytic cycle of a bacteriophage infection. In this process, bacteriophages infect bacterial cells, replicate within them, and ultimately cause cell lysis, releasing newly assembled virions. Occasionally, random fragments of the bacterial genome...
2.6K
Mechanism of Conjugation
1.5K
Bacterial conjugation is a mechanism of horizontal gene transfer that enables the exchange of genetic material between bacterial cells through direct contact. This process is facilitated by a donor cell carrying a conjugative plasmid, which encodes genes necessary for pilus formation, DNA replication, and transfer. The conjugative plasmid plays a central role in initiating and executing the transfer of genetic material.The tra region of the conjugative plasmid encodes proteins responsible for...
1.5K
Transposons
2.9K
Transposons, or "jumping genes," are small mobile genetic elements (MGEs) that range from 700 to 40,000 base pairs in length. They are found in all organisms and can move within the same chromosome or transfer to different chromosomes. In some cases, transposons can also jump between different host DNA molecules, such as plasmids or viruses, contributing to genetic variability.Barbara McClintock first discovered these mobile genetic elements in the 1940s while studying maize genetics, and she...
2.9K
DNA-only Transposons
18.5K
DNA-only transposons are called autonomous transposons since they code for the enzyme transposase that is required for the transposition mechanism. Insertion of transposons can alter gene functions in multiple ways. They can mutate the gene, alter gene expression by introducing a novel promoter or insulator sequence, introduce new splice sites, and change the mRNA transcripts produced, or remodel chromatin structure.
The donor site from where the transposon is excised is either degraded or...
The donor site from where the transposon is excised is either degraded or...
18.5K
Overview of Transposition and Recombination
20.6K
Transposons make up a significant part of genomes of various organisms. Therefore, it is believed that transposition played a major evolutionary role in speciation by changing genome sizes and modifying gene expression patterns. For example, in bacteria, transposition can lead to conferring antibiotic resistance. Movement of transposable elements within the genetic pool of pathogenic bacteria can aid in transfer of antibiotic-resistant genetic elements. In eukaryotes, transposons can carry out...
20.6K
Translesion DNA Polymerases
11.8K
Translesion (TLS) polymerases rescue stalled DNA polymerases at sites of damaged bases by replacing the replicative polymerase and installing a nucleotide across the damaged site. Doing so, TLS allows additional time for the cell to repair the damage before resuming regular DNA replication.
TLS polymerases are found in all three domains of life - archaea, bacteria, and eukaryotes. Of the different classes of TLS polymerases, members of the Y family are fitted with specialized structures that...
TLS polymerases are found in all three domains of life - archaea, bacteria, and eukaryotes. Of the different classes of TLS polymerases, members of the Y family are fitted with specialized structures that...
11.8K
