RNASEH1 E.Coli

Ribonuclease H1 E.Coli Recombinant
Cat. No.
BT26664
Source
Escherichia Coli.
Synonyms
Ribonuclease HI, RNase HI, Ribonuclease H, RNase H, rnhA, dasF, herA, rnh, sdrA, b0214, JW0204.
Appearance
Sterile Filtered clear solution.
Purity
Greater than 95% as determined by SDS-PAGE.
Usage
THE BioTeks products are furnished for LABORATORY RESEARCH USE ONLY. The product may not be used as drugs, agricultural or pesticidal products, food additives or household chemicals.
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Description

RNASEH1 E.coli Recombinant produced in E.Coli is a single, non-glycosylated polypeptide chain containing 178 amino acids (1-155 a.a.) and having a molecular mass of 20kDa.
RNASEH1 is fused to a 23 amino acid His-tag at N-terminus & purified by proprietary chromatographic techniques.

Product Specs

Introduction
RNase H is an enzyme that specifically breaks down the RNA strand in RNA:DNA hybrids. It is found in the nucleus, where it plays a critical role in DNA replication by removing the RNA primers from Okazaki fragments on the lagging strand. RNase H cleaves RNA in the middle of the strand, leaving a 5' phosphate group. It requires either magnesium or manganese ions for activity.
Description
Recombinant E. coli RNase H1 is a non-glycosylated protein with a molecular weight of 20 kDa. It is produced in E. coli and consists of a single polypeptide chain of 178 amino acids (residues 1-155), with a 23-amino acid His-tag fused to the N-terminus. The protein is purified using proprietary chromatographic techniques.
Physical Appearance
Clear, sterile-filtered solution.
Formulation
The RNase H1 protein solution has a concentration of 1 mg/ml and is supplied in a buffer containing 20 mM Tris-HCl (pH 8.0), 10% glycerol, and 2 mM DTT.
Stability
For short-term storage (2-4 weeks), the product can be stored at 4°C. For longer storage, it is recommended to freeze the product at -20°C. Adding a carrier protein like HSA or BSA to a final concentration of 0.1% is recommended for long-term storage. Avoid repeated freeze-thaw cycles.
Purity
The purity of the RNase H1 protein is greater than 95% as determined by SDS-PAGE.
Synonyms
Ribonuclease HI, RNase HI, Ribonuclease H, RNase H, rnhA, dasF, herA, rnh, sdrA, b0214, JW0204.
Source
Escherichia Coli.
Amino Acid Sequence
MGSSHHHHHH SSGLVPRGSH MGSMLKQVEI FTDGSCLGNP GPGGYGAILR YRGREKTFSA GYTRTTNNRM ELMAAIVALE ALKEHCEVIL STDSQYVRQG ITQWIHNWKK RGWKTADKKP VKNVDLWQRL DAALGQHQIK WEWVKGHAGH PENERCDELA RAAAMNPTLE DTGYQVEV.

Product Science Overview

Introduction

Ribonuclease H1 (RNase H1) is an essential enzyme involved in the maintenance and replication of DNA. It specifically degrades the RNA strand of RNA-DNA hybrids, which are formed during various cellular processes such as DNA replication and repair. The recombinant form of RNase H1 from Escherichia coli (E. coli) is widely used in research due to its high activity and stability.

Structure and Function

RNase H1 belongs to the RNase H family of enzymes, which are characterized by their ability to cleave the RNA strand of RNA-DNA hybrids. The enzyme consists of a single polypeptide chain with a molecular weight of approximately 18 kDa. It contains a conserved catalytic domain that is responsible for its enzymatic activity .

The primary function of RNase H1 is to remove RNA primers from Okazaki fragments during lagging strand DNA synthesis. This activity is crucial for the completion of DNA replication and the maintenance of genomic stability. Additionally, RNase H1 plays a role in the processing of R-loops, which are RNA-DNA hybrid structures that can form during transcription .

Recombinant Expression in E. coli

The recombinant expression of RNase H1 in E. coli involves the insertion of the RNase H1 gene into a suitable expression vector, which is then introduced into E. coli cells. The bacteria are cultured under conditions that promote the expression of the recombinant protein. The expressed protein is then purified using various chromatographic techniques to obtain a highly pure and active enzyme .

Applications in Research

Recombinant RNase H1 from E. coli is widely used in molecular biology research. Some of its key applications include:

  • DNA Replication Studies: RNase H1 is used to study the mechanisms of DNA replication, particularly the removal of RNA primers and the processing of Okazaki fragments.
  • R-loop Analysis: The enzyme is employed to investigate the formation and resolution of R-loops, which are important for understanding transcriptional regulation and genome stability.
  • Antisense Oligonucleotide Research: RNase H1 is used to evaluate the efficacy of antisense oligonucleotides, which are designed to bind to specific RNA sequences and induce their degradation .

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