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

Describe the plasma homocysteine levels, Describe the Plasma Homocysteine l...

Describe the Plasma Homocysteine levels ? Several studies have investigated the contribution of homocysteine to CVD risk both among immigrant Indians and those living in India.

Define procedure for determination of fungal and yeast count, Define Proced...

Define Procedure for Determination of Fungal and Yeast Count? Now carry out the exercise following the steps enumerated herewith: 1. Shake the food sample and aseptically tr

Bovine viral diarrhoea, B o v i ne viral diarrhoea Bovine viral dia...

B o v i ne viral diarrhoea Bovine viral diarrhoea (BVD) and mucosal disease (MD) are clinically dissimilar disease syndrome yet have a common viral etiology. The acute dise

Explain the psychological management of eating disorders, Explain the Psych...

Explain the Psychological Management of eating disorders? All anorexia nervosa or bulimia nervosa patients are resistant to any kind of therapy and hospitalization may be a lif

Fertilization - pre-embryonic development, Fertilization - Pre-Embryonic De...

Fertilization - Pre-Embryonic Development Ovulation generally takes place between 9th to 14th day after the end of last menstrual period. Fertilization of oocyte initiates dev

Human factor theory - accident causation, Human Factor Theory - Accident Ca...

Human Factor Theory - Accident Causation This theory of accident causation suggests that a chain of events caused by human error is the cause of accident. The chain of events

Explain the aseptic technique - sub culturing, Explain the Aseptic Techniqu...

Explain the Aseptic Technique - Sub Culturing? Aseptic technique is a series of procedures used to prevent contamination of cultures and sterile culture media. It is one of the

Microscopic examination of sputum, Question 1 Discuss briefly the collecti...

Question 1 Discuss briefly the collection and examination of pericardial fluid Question 2 Discuss the following Collection of CSF Physical examination of CSF

Describe the various steps involved in glycolysis, Question 1 Give a detai...

Question 1 Give a detailed account on "high energy compounds" Question 2 Describe the various steps involved in Glycolysis Question 3 Give a detailed account on Oxidativ

What is metabolism , What is Metabolism ? Metabolism is usually define...

What is Metabolism ? Metabolism is usually defined as the sum of the chemical processes required to sustain life. These processes, called metabolic pathways, are catalyzed by

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