Determine cdna encodes aba 8-hydroxylase activity, Biology

Assignment Help:

Hypothetical question. A new biotech company intends to clone the gene encoding the ABA 8-hydroylase enzyme, an important protein regulating plant hormone degradation that controls seed dormancy and environmental stress responses. The protein is known to belong to a class of enzymes known as p450s of which there are 500 different genes in Arabidopsis, the model plant system. The company isolates a full length cDNA for each gene using PCR with primerscontaining attB sequences and then each individually cloned into pDONR/Zeo (from Invitrogen) via a BP reaction for all 500 different p450 genes from Arabidopsis (the cDNAs vary in length from 1.5-2.5 kb). Next the company proposes to express each gene in yeast, and then carry out an ABA-8 hydroxylase reaction in vivo to determine which cDNA encodes ABA 8-hydroxylase activity.

Outline a cloning strategy(s) (experimental procedure) the company could take to express all these genes in yeast - a simple flow diagram detailing the steps is all that is needed. Design a vector(s) that would be central in this strategy (can just draw by hand) and list all the important features that would be required (i.e what cloning sites or antibiotic resistance genes would be required if any) but not any features that would be excess to what is required.

 


Related Discussions:- Determine cdna encodes aba 8-hydroxylase activity

Corelation of st depressoin with coronary angiography, Q. Corelation of ST ...

Q. Corelation of ST Depressoin with Coronary Angiography? Various investigators considered 1 mm of horizontal or downsloping ST depression to denote a positive test and used fr

Explain basic functions of the micronucleus, Q. What are the respective fun...

Q. What are the respective functions of the micronucleus and of the macronucleus in the paramecium? The macronucleus is properly the cell nucleus it has RNA and DNA and acts as

Explain expansion in red cells during pregnancy, Explain expansion in Red C...

Explain expansion in Red Cells during pregnancy? Production of red cells is stimulated and their numbers gradually rise.  However, as compared to the expansion in plasma volume

Risk factors of cardiovascular disease, Q. Show Risk factors of cardiovascu...

Q. Show Risk factors of cardiovascular disease? Various risk factors of cardiovascular disease. We will now briefly mention the causes related to hypertension. 1. Genetic

Internal budget - nutrient budgets, Internal budget - Nutrient Budgets ...

Internal budget - Nutrient Budgets This is concerned with the circulation of nutrients through various biotic and abiotic compartments of a given ecosystem. In other words the

Define nutritional requirement in cold and polar environment, Define Nutrit...

Define Nutritional Requirements in Cold and Polar Environment? Energy requirements are the major consideration for providing nutritional support in a cold environment. Energy e

Determine the seeds in the plant kingdom, What is the evolutionary importan...

What is the evolutionary importance of the emergence of seeds in the plant kingdom? The evolutionary significance of the seed is related to the plant capability of distant colo

Discuss the example of comprehensive neuropsychology, Discuss the example o...

Discuss the example of Comprehensive neuropsychology  Example of these include deficits of visuo-spatial perception hampering performance on construction tasks, poor comprehe

Write Your Message!

Captcha
Free Assignment Quote

Assured A++ Grade

Get guaranteed satisfaction & time on delivery in every assignment order you paid with us! We ensure premium quality solution document along with free turntin report!

All rights reserved! Copyrights ©2019-2020 ExpertsMind IT Educational Pvt Ltd