Plasmid Extraction

This essay has a total of 400 words and 3 pages.

Plasmid Extraction


Introduction

Chitobiase, from Vibrio harveyi, is a membrane bound lipoprotein involved in the
degradation of chitin. Chitobiase is similar to and may share a common ancestry to the
a-chain of human b-hexos-aminidase. Chitobiase is encoded by chb.


In this experiment, a restriction map for restriction enzymes Eco R1, Pst1 and Hind III
using Southern hybridization and restriction analysis of pRSG 192. pRSG 192 is a
recombinant plasmid derived from the chb gene and pUC 19, a 2.7kb engineered plasmid which
encodes for ampicillin resistance, a portion of the lac operon and a multiple cloning
region . The chb gene exists as a 3.6 kb insert in the mutiple cloning region of pUC 19.


The major goals of Experiment One will be to isolate pRSG 192 from an overnight culture of
E. coli, amplify a region of the chb gene using PCR, and to map restriction sites within
the chb gene using restriction analysis and Southern hybridization.


Methods
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