Transformation Experimental Design (will be collected and graded by yourTA) Assu

Question

Transformation Experimental Design (will be collected and graded by yourTA) Assuming you work in a pharmaceutical company. You are the lead scientistin charge of developing a research project to genetically transform bacteria toproduce humaninsulin. If your research is successful m ons of diabetics will be able to use the syntheticinsulin your company produced to regulate their bloodsugar levels. coli Beta-galactosidas rols the the insulin gene. For E coli to produce insulin, you are given a genetically engineered strain that contains recombinant human insulin, B-galactosidase, and tetracycline (an antibiotic) resistance genes. There are two strains of E coli bacteria (strain A and B) known to uptake this plasmid easily. You would like to know which strain of E coli is easier to be genetically transformed, i e. has higher efficiency in producing insulin. Design a simple experiment for your research. brief protocol for the manufacturing division in your company. In this protocol, you don't have to include all the detailed experimental procedures (such as pg. 1 and 2). However, you should clearly describe: 1) Materials required to conduct this experiment. 2) The control and the experimental treatments. 3) Instruction on how you would set up the treatments. 4) How do you select which strain of E coli is more suitable for your insulin production? Write the protocol in a piece of paper and submit it to your TA.TA may deduct points on incomplete protocol or poorly designed experiment.

Explanation / Answer

4) to find out the which strain is more best we can calculate the transformation efficiency of both the strains of the bacteria after transformation and plating.. The strain which is giving the more colonies on the plate is more efficient in transformation..!!

3) as we have been given a Plasmid which is having a tetracycline resistance gene.. The protocol will be.

(a) perform transformation at 42°C with both the strains..

(b) put the Autoclave LB broth in the transformed tube. Keep I for 1 hour a 37°C..

(c) preapre the L.B + Tetracycline antibiotics plates for each strains..

(d) spread some amount of the incubated transformation E.coli culture on the plates separately..

(e) observe for the colonies next day..!!

Only transformed bacteria will grow on the antibiotic plates..!!

1) the materials will be :-

(a) plates

(b) L.B

(c) tetracycline

(d) bacterial strain

(e) recombinant Plasmid..

2) negative control - untransformed bacterial strains on the tetracycline plates..!!!

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