広域宿主プラズミドRK2の分子媒介特性

R Meyer, D Figurski, D R Helinski

Science (New York, N.Y.)

|December 19, 1975

まとめ

プラズミドRK2は,多くの細菌で安定的に維持されます. DNAをEco RIサイトに挿入すると,プラズミドの機能に影響を及ぼさず,新しいハイブリッドプラズミドを作成するのに理想的です.

科学分野:

分子生物学は分子生物学である.
微生物学微生物学とは
遺伝学遺伝学とは

背景:

プラズミドRK2は,様々なグラム陰性細菌の安定維持のために知られている.
プラズミドの安定性と移転を理解することは,遺伝子工学にとって極めて重要です.

研究の目的:

RK2プラズミドのユニークなEco RIサイトにDNA挿入の機能的影響を調査する.
グラム陰性細菌におけるハイブリッドプラズミドを構成するためのベクターとしてのRK2の有用性を評価する.

主な方法:

プラズミドDNA操作は,RK2のEco RIサイトにDNA断片を挿入することを含む.
プラズミドの安定性と自己伝染性の評価を,様々なグラム陰性細菌宿主において行いました.

主要な成果:

エコ・リ・サイトにDNA断片を挿入しても,プラズミドの維持は妨げられませんでした.
改造されたRK2プラズミドの自己感染性は影響を受けませんでした.
RK2は,広範囲のグラム陰性細菌の宿主の中で安定した維持を示した.

結論:

RK2プラズミドは,グラム陰性細菌の遺伝子操作のための多用途で堅牢なベクターです.
ユニークなEco RIサイトは,ハイブリッドプラズミドのin vitro構築に適した場所を提供します.
RK2ベースのハイブリッドプラズミドは,結合または変換を通じて,さまざまなグラム陰性生物に容易に確立することができます.

さらに関連する動画

11:19

Genome-wide Analysis using ChIP to Identify Isoform-specific Gene Targets

Published on: July 7, 2010

12:48

Measuring Cation Transport by Na,K- and H,K-ATPase in Xenopus Oocytes by Atomic Absorption Spectrophotometry: An Alternative to Radioisotope Assays

Published on: February 19, 2013

関連する概念動画

02:36

RNA Polymerase II Accessory Proteins

Proteins that regulate transcription can do so either via direct contact with RNA Polymerase or through indirect interactions facilitated by adaptors, mediators, histone-modifying proteins, and nucleosome remodelers. Direct interactions to activate transcription is seen in bacteria as well as in some eukaryotic genes. In these cases, upstream activation sequences are adjacent to the promoters, and the activator proteins interact directly with the transcriptional machinery. For example, in...

02:30

Nucleophiles

The word “nucleophile” has a Greek root and translates to nucleus-loving. Nucleophiles are either negatively charged or neutral species with a pair of electrons in a high-energy occupied molecular orbital (HOMO). As these species tend to donate electron pairs, nucleophiles are considered Lewis bases as well. Negatively charged species, like OH−, Cl−, or HS−, with one or several pairs of electrons, are typically nucleophiles. Similarly, neutral species such as ammonia, amines, water, and alcohol...

01:42

The ADP/ATP Carrier Protein

ADP/ATP carrier or AAC protein is the most abundant carrier protein in the inner mitochondrial membrane. It transports large quantities of ADP and ATP, equivalent to the average human body weight, every day. Among other transporters, ACC protein is one of the best-studied members of the mitochondrial carrier protein family. The ADP/ATP carrier protein comprises two transmembrane helices connected to a loop and a single alpha-helix on the matrix side. It switches between two conformational...

01:14

Rab Proteins

Rab proteins constitute the largest family of monomeric GTPases, of which 70 members are present in humans. Rab proteins and their effectors regulate consecutive stages of vesicle transport such as vesicle transport, docking, and fusion to the correct recipient membrane.
Rab proteins switch between a cytosolic, GDP-bound inactive state and a membrane-anchored, GTP-bound active state. By themselves, Rabs show slow rates of GDP/GTP exchange and GTP hydrolysis. Thus, Rab proteins are considered...

02:36

RNA Polymerase II Accessory Proteins

01:26

Receptor Tyrosine Kinases

Receptor tyrosine kinases or RTKs are membrane-bound receptors that phosphorylate specific tyrosine on protein substrates. RTKs regulate cellular growth, differentiation, survival, and migration. They contain an extracellular ligand binding domain, a transmembrane domain, and a cytosolic tail with intrinsic kinase activity. Several extracellular signaling molecules activate RTKs in one or more ways and relay the signal downstream. Ligands such as platelet-derived growth factor (PDGF) or...

関連する実験動画