How does blue white screening work? What gene is required? What is the name of t

Question

How does blue white screening work? What gene is required? What is the name of the protein that it encodes? What is X-gal? What does it allow you to distinguish about a cloning and transformation reaction? What types of genes or DNA sequences are required for a plasmid vector? Assume you ligate a target DNA sequence to a linearized cloning vector (pUC18/19) that is cut within its lacZ gene... i. What DNAs produced from the ligation reaction would successfully transform bacteria? Draw some of the possible products of ligation. Ii. Which of the products that you drew would result in colorless colonies if you transformed bacteria with it?

Explanation / Answer

The answers
1. Blue white screening is working on based on the product of selection gene. If the gene product is functional (non transforment) the colony will appear in blue in colour, if the gene product is non functional (if transforment) the colony will apper in white in colour.

For Blue white screening LacZ gene is required.

It encodes for -galactosidase enzyme.

5-bromo-4-chloro-3-indolyl--D-galactopyranoside cyclohexa ammounium salt is called as X gal. It has Indole which is linked to Indole.

The functional product of lacZ gene can react with X gal and gives blue colour, where as non functional product of lacZ gene can not react with X gal which leads to white colour. Using this principle this technique is employed.

2.
For a plasmid vector it needs,
a replication gene - ori gene,
selection marker - antibiotic,
MCS/PLS site - for gene insertion and
Reporter gene - either antibiotic or non antibiotic.

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