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

Define influence of gardening on the development of botany, Define Influenc...

Define Influence of Gardening on the Development of Botany? The influence of gardening on the development of botany may be seen in the work of John Gerard (1545-1612). Gerard p

Spirogyra, whats the meaning of asexual reproduction

whats the meaning of asexual reproduction

Explain phylum phaeophyta, Phylum Phaeophyta (brown algae) 1) They are ...

Phylum Phaeophyta (brown algae) 1) They are brown algae, apart from chlorophyll they have brown pigment i.e. fucoxanthin which predominates. 2) They are one of ,the most com

What is composition, What is Composition? Blood consists of red blood c...

What is Composition? Blood consists of red blood cells, white blood cells, and platelets, suspended in a straw-colored liquid called plasma. If whole blood is centrifuged, abou

Questions, assignment having 12 questions

assignment having 12 questions

Determine the general principles for incisions, General Principles for Inci...

General Principles for Incisions The general principles for placing incisions so as to create surgical flaps remain the same regardless of the reason for creating them. These a

Explain amylopectin, Amylopectin Amylopectin forms a branched structure...

Amylopectin Amylopectin forms a branched structure with about 30 glucose units in a chain among branches. There are usually slightly more  'outer' unbranched chains (called A-c

Cellular level - level of body organization, Cellular level - Level of body...

Cellular level - Level of body organization The next higher level of organization is the cellular level. This is really an aggregation of cells that are functionally different

Briefly explain about phase of the plasmodium life cycle, Q. To which phase...

Q. To which phase of the plasmodium life cycle do the typical chills and fever of malaria correspond? The typical fever and chills episodes of malaria correspond to the phase w

Adverse effects of air pollution, Air pollution affects both living and non...

Air pollution affects both living and non-living matter. The effects can be classified as.

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