Abstract
The advent of recombinant DNA technology has brought a series of dramatic changes in biology.
advent 출현
It offered new opportunities for innovations to produce a wide range of therapeutic products. Recombinant DNA technology refers a series of procedures used to produce recombinant DNA (rDNA) molecules.
innovation 혁신
therapeutic 치료상의
procedures 절차
The first step in recombinant DNA technology is to select a piece of DNA to be inserted into a vector.
The second step is to cut that piece of DNA with a restriction enzyme and then ligate the DNA insert into the vector with DNA Ligase. And the third step is inserting the combination into host cells.
ligate 묶다
DNA ligase is an enzyme that catalyzes the formation of a covalent bond between a 5' phosphate and a 3' hydroxyl group at the termini of DNA fragments.
A vector is a small piece of DNA molecule, which is used as a vehicle to artificially carry foreign genetic material into host cell, where it can be replicated and/or expressed.
vehicle 수단
artificially 인위적인
replicate 복제하다
express 나타내다
Vectors have three main features: origin of replication, multiple cloning sites and selectable marker. Cloning vector and expression vector are two types of vectors, used in recombinant DNA technology.
A cloning vector is a small piece of DNA used to introduce genes into cells while obtaining numerous copies of the insert.
obtaining 취득
Vectors used for cloning include: plasmids, bacteriophages, cosmids, bacterial artificial chromosomes (BACs), yeast artificial chromosomes (YACs) and human artificial chromosomes (HACs).
Expression vectors are mainly plasmids, designed for the transcription and protein expression of the transgene. Since all vectors had been created to meet different purposes, each cloning vector and expression vector should clearly be described.
transcription
transgene 이식유전자
Introduction
The advent of recombinant Deoxy Ribonucleic Acid (DNA) technology revolutionized the development in biology, and led to a series of dramatic changes.
advent 출현
revolutionize 대변을 일으키다
It offered new opportunities for innovations to produce a wide range of therapeutic products with immediate effects (Li et al., 2017).
innovation 혁신
Recombinant DNA technology is a science by which series of procedures are used to produce recombinant DNA (rDNA) molecule, a piece of DNA that has been created by the combination of at least two strands.
procedures 절차
It is possible because DNA molecules from all organisms share the same chemical structure, and differ only in the nucleotide sequence within that identical overall structure [21] .
identical overall structure 동일한 전체 구조
The first step in recombinant DNA technology is to select a piece of DNA to be inserted into a vector.
The second step is to cut that piece of DNA with a restriction enzyme and then ligate the DNA insert into the vector with DNA Ligase, and third step is inserting the combination into host cells. A vector is a small piece of DNA molecule, which is used as a vehicle to artificially carry foreign genetic material into host cell, where it can be replicated and/or expressed.
ligate 묶다
artificially 인공적으로
A vector containing foreign DNA is termed as recombinant DNA. The purpose of a vector is typically to isolate, multiply, or express the insert in the target cell [16] .
Useful vectors have three main features: (i) An origin of replication (Ori), which is a specific DNA sequence at which DNA replication is initiated.
The essential feature of a vector is that it can replicate autonomously in a host species, usually bacteria. The origin is therefore absolutely essential for the amplification of the vector inside a bacterial host.
replicate 복제하다
autonomously 독자적으로
amplification 증폭
inside 안에
(ii) Presence of a multiple cloning site (MCS), a region with multiple useful restriction enzyme sites to make a compatible digest of the vector and DNA fragment.
multiple 많은
restriction enzyme 제한효소
compatible 양립될 수 있는
digest 소화
(iii) A selectable marker, which is a method of allowing hosts (usually bacteria) containing the vector to be readily identified and purified.
purify 정제[제련]
The insert contains a selectable marker which allows for identification of recombinant molecules.
An antibiotic marker is often used so that a host cell without a vector dies when exposed to a certain antibiotic, and the host with the vector will live because it is resistant [13] .
Cloning Vectors
Gene cloning is a major breakthrough, the important part of which is a cloning vector.
breakthrough 돌파구
The various uses of cloning remain redundant if a suitable cloning vector is not chosen.
redundant 불필요한, 쓸모없는
A cloning vector is a fraction of DNA that can be used to insert a foreign DNA molecule and has the ability to be inserted into a host for cloning purpose [8].
fraction 부분
It is capable of self-replication inside the host cell. The purpose of a cloning site is to provide a place for cloning to occur. A selectable marker gene helps identifying successful recombinants after cloning.
The cloning vector does not necessarily help to express a protein which the foreign DNA encodes.
Thus, the sole purpose of the cloning vector is to carry foreign DNA to the host. Depending on the size and the application of the insert the suitable vector is selected for a particular purpose [14] .