Question

Outline the roles played by restriction enzymes and vectors in cloning DNA.

Words to choose from: bacterial, eukaryotic, nucleotide, DNA, RNA, protein, palindrome,

nuclease, protease, phage, plasmid.

1. A restriction enzyme is a _________      ____________that recognizes specific _________ sequences in a ________molecule, often a __________, and cleaves or nicks the molecule at those sites.

2. A vector is an agent into which a foreign DNA segment is inserted and used to transform host cells. A vector can be created from a virus known as a __________ or a bacterial _________.

Answer 1

Cloning is used for amplification and insertion of gene of interest in expression vector for getting protein in large in large amount.

In cloning every component is important from vector , gene of interest , expression cell line, restriction enzyme,primers etc.

Restriction enzyme: restriction enzymes are the enzymes which cleaved dsDNA by recognising specific nucleotides on DNA ,so these enzymes are used for making complementary binding sites on vector and GOI(gene of interest). We select RE (restriction endonuclease) by looking in our GOI And vector .2 RE are selected .the RE we selected should not cut withing the GOI and our vector should not have more then one RE site and these sites should be in MCS near promoter.these things should be made sure before cloning.

Vectors are also important because they have promoter system ,anybody resistant gene ,MCS etc.

There are many vector such ase plasmid ,cosmid ,phagemid, bacterial artificial vector, yeast artificial vectors etc.

The main function of vector is to provide promoter system for the large scale expression of protein from gene of interest.it also selects the cells by providing antibiotic resistance.

Fill in the blanks

1.a. nuclease

b. nucleotide

c. DNA

d. palindrome

2. a. phage

b. plasmid