Determine cdna encodes aba 8-hydroxylase activity, Biology

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.

 

Posted Date: 3/11/2013 1:34:21 AM | Location : United States







Related Discussions:- Determine cdna encodes aba 8-hydroxylase activity, Assignment Help, Ask Question on Determine cdna encodes aba 8-hydroxylase activity, Get Answer, Expert's Help, Determine cdna encodes aba 8-hydroxylase activity Discussions

Write discussion on Determine cdna encodes aba 8-hydroxylase activity
Your posts are moderated
Related Questions
Phylum Mollusca (47,000 species) The segmentation and coelom may not exist. Diversification through modifications of head, foot; mantle and 'radula for feeding. .Visceralorgans

Slight or no Metamorphosis In primitive wingless insects (Apterygota) such as spring tails, silver fish etc. and in secondarily apterous insects the young ones which hatch fr

Define Use of Isotopically Labelled Nutrients? Nutrient Turnover Radioactive labelled nutrients are used to know the total body pool and the compartment in which it is stored.

Pre-surgical Prosthodontics Radiographic investigation: The radiologic investigations are critical to evaluate the bone quality and bone quantity. 1. Panoramic Radiograph

Q. What are flagella and cilia and how do these structures acquire movement? What are some examples of flagellated and ciliated cells in humans? Flagella and Cilia are structur

Mode of nutrition of a rabbit

TYPES OF SPERM Mainly two types . (1) Flagellated Sperm - Generally all sperm are monoflagelated, but in todfish or opsonus sperm is biflagelated. (2) Non flagellated spe

1.   Carbon monoxide (CO): It is colourless, odourless, tasteless gas and is not soluble in water. Source: CO is produced due to: (i)     Incomplete combustion of fuels

what characteristics of fatty acids are false? (Dissolves in water, used for energy storage, used to build lipids, found in triglycerides, found in steroids)?

Embryos Prom Synergids The synergids, which usually degenerate prior to or soon after double fertilization, are reported to give rise to embryos in Argentine mexicana, Tamari