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
You are working at a large, well funded research institute with the latest biochemical techniques available. The focus of your research is to find a specific mutation, or set of mu

Indirect determination: By determining the weight of the filler on the dried filter paper you can calculate the weight of aspirin in each tablet: weight of acetylsalicylic ac

Sarcodine protozoans are amoebas, and there are two types of them: shelled and naked. As you examine the prepared slides you should be able to see water expulsion vesicle, nucleus,

Q. Is there an exchange of cells between the mother and the fetus through the placenta? Under usual conditions there is no passage of cells across the placenta during gestation

Define Requirements of Fluid in Postoperative Nutritional Care Extensive fluid losses may occur through vomiting, haemorrhage, diesis, excudate, fever and sweating after a surg

Heifers As efficient reproductive performance is essential for economic livestock production, the female calves must grow rapidly to attain sexual maturity, ovulate and be mated b

Imiquimod (Aldara) This immunomodulator is FDA-approved for topical treatment of external and perianal genital warts, which are caused by papillomavirus. Gradual clearance of w

Problem 1: Show the aims and applications of Epidemiology. Definition of Epidemiology. List aims of Epidemiology List applicationsof Epidemiology Problem 2

RECAPITUL A TIO N THEORY / BIOGENETIC LAW It was proposed by Muller & Haeckel on the basis of Baer's law. According to it, in the development of animal phylum characters

Illustrate the types of Bone Remodeling Bone remodeling occurs on existing bone surfaces. However, unlike modeling, remodeling cannot cause large changes in bone structure at a