Describe one technique you would use to study whether the recombinant GATA-1 pro
ID: 3166924 • Letter: D
Question
Describe one technique you would use to study whether the recombinant GATA-1 protein made and purified from bacteria can recognize its target enhancer sequence on the beta amylase upstream region. (3 points) Wesien loting b.) Describe a technique compared to a breast cancer tissue. (3 points) for studying the entire transcriptome of a normal breast tissue when vevcheransoviptasé Pce c.) Explain the difference between gene addition and gene replacement. (2 points) lar sit in a d.) What is the purpose of creating a gene knockout animal? (1 point) e.) Give two reasons it may be advantageous to produce human proteins by recombinant DNA technology in a cukaryotic cell such as fungi than in bacteria. (2 points) 11 points 3Explanation / Answer
a. Electrophoretic Mobility Shift Assay (EMSA)
Incubate the recombinant GATA-1 protein with the enhancer sequence and run them in a native PAGE gel. If GATA-1 binds to the enhancer sequence, we can observe a supershift in the banding pattern.
b. To study entire transcriptome of a tissue, we can perform
i. Microarray analysis: RNA isolated from the tissue is converted to cDNA and probed to a small chip that contains complementary sequences
ii. RNA sequencing: Whole RNA sequences are counted and sequenced
c. Gene addition involves the addition of a new gene to an existing set of chromosomes. Gene replacement involves replacement of an existing gene with a new gene.
d. Gene knock out involves removal of the genomic sequence of a particular gene from the genome. Gene knockouts are used to study the loss of function phenotypes of genes and their effect on physiology or development.
e.
i. High amounts of production
ii. Rapid production
iii. Low cost