A group of organic molecules are called Proteins. Proteins are very essential of all the materials that make up all living organisms. Proteins are chains of amino acids, programmed by DNA. Proteins are copious in different types and are present in every single living cell of all organisms. The uniqueness of the organisms are found out by protein activity for instance, eye color, immune system operation, etc,. Recombinant protein is a protein that whose code is carried by a recombinant DNA. The term recombinant DNA means that two segments of DNA in a plasmid. Plasmids are those which generally occur in bacteria. Once a recombinant DNA is inserted into bacteria, these bacteria will make protein based on this recombinant DNA. This protein is known as "Recombinant protein". Now-a-days, the utilization of bacteria for producing recombinant protein is very advanced. Recombinant DNA (rDNA) molecules are DNA sequences that is a consequence of molecular cloning (used to assemble recombinant DNA molecules) to assemble the genetic material from various sources, making sequences that cannot be predicted in biological organisms. This technique is often used to produce human growth hormone and insulin. Occasionally, Recombinant DNA molecules are known as “chimeric DNA”, as they are made of material from two species.
The method of recombinant DNA was initially planned by a graduate student, Peter Lobban, along with a biochemist, A. Dale Kaiser at the Stanford University. During then years,1972–74, the method was then acknowledged by Stanley Norman Cohen, an American geneticist Chang, Herbert Boyer, a addressee of the 1990 National Medal of Science. In 1973, they published their predictions in journal "Enzymatic end-to-end joining of DNA molecules" which explained the methods to separate and intensify genes or DNA segments and introduce them into an additional cell with accuracy. In 1977, an advance in the field of recombinant DNA technology took place when Herbert Boyer created the biosynthetic "human" insulin, a group of biosynthetic human insulin products.
The DNA sequences that are utilized in the recombinant DNA molecules building can initiate from any species. For instance, plant DNA may be combined to bacterial DNA, or human DNA may be associated with fungal DNA. Also, DNA sequences that may not transpire anywhere in environment may be produced by the chemical synthesis of DNA, and built-in into recombinant molecules. Any DNA sequence may be created and introduced into any of a very wide range of living organisms, with the help of artificial DNA and recombinant DNA method. Recombinant proteins are those which is an outcome of the expression of recombinant DNA inside living cells. Once the recombinant DNA encoding a protein is set up into a host organism, the creation of recombinant protein is not required.
The genes that code for the proteins are put into special vectors, or units of DNA. Vectors are selected that will create huge quantities of the preferred protein. This is known as “over expression”. Over expression is performed in unique host cells. Occasionally, the hosts are bacteria or yeast. The hosts are frequently insect or mammalian cell lines, in cases where the proteins are from mammals. Various kits are commercially present to assist both the cloning of the gene, and the following recombinant protein fabrication.
There are basically two methods for producing recombinant protein.
• One is the molecular cloning, a laboratory method used to make recombinant DNA.
• The other method is the polymerase chain reaction (a method in molecular biology to intensify a single or a few copies of a piece of DNA) used to proceed the replication of any specific DNA sequence selected by the scientist.
The basic difference between the two methods is that molecular cloning incorporates the replication of the DNA within a living cell, whereas PCR replicates DNA in the test tube, without living cells. DNA replication is the process of making an identical copy of a section of duplex (double-stranded) DNA, using existing DNA. Configuration of recombinant DNA needs a cloning vector, a DNA molecule that will replicate inside a living cell. Generally, vectors are a consequent of plasmids or viruses, and stand for fairly small segments of DNA that contain necessary genetic signals for replication. The option of vector for molecular cloning relies on the option of host organism, the size of the DNA to be cloned, and the way in which the foreign DNA is to be expressed.
The typical cloning etiquette any DNA fragment fundamentally involves seven steps: