Chapter 18.2, Problem 2R

Summary Introduction

To review:

The sequential steps for inserting a foreign DNA (deoxyribonucleic acid) into a plasmid or vector by arranging the below-mentioned steps in a correct order:

a. Transform host cells.

b. Select colonies for antibiotic resistance.

c. Select colonies that do not grow under ultraviolet light.

d. Digest vector and foreign DNA with a restriction enzyme.

e. Ligate the digested plasmid together with the foreign DNA.

Introduction:

Plasmids are small, extra-chromosomal DNA molecules. They are usually circular and are relatively smaller (2000–6000 base pairs). There are often multiple restriction sites present in a plasmid sequence. Thus, this makes it easier for the insertion of a new foreign DNA. In the given case, it is assumed that a plasmid vector contains genes for green fluorescent protein (GFP) as well as for ampicillin resistance. A restriction site is present on the gene for GFP only. A foreign DNA needs to be inserted into this plasmid, which in turn, is to be inserted into the bacteria.