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

What is crossing over, What is crossing over? In which period of meiosis do...

What is crossing over? In which period of meiosis does this event occur? Crossing over is the eventual exchange of chromosomal fragments among homologous chromosomes. The pheno

What is the main theory opposed to evolution, What is the main theory oppos...

What is the main theory opposed to evolution? The major theory that opposes the evolution theory on the description of how species emerged (phylogenesis) is fixism.

Investigation of aortic regurgitation by exercise testing, Q. Investigation...

Q. Investigation of aortic regurgitation by Exercise Testing? It is helpful in assessing functional capacity, symptom status and hemodynamic effects in patients with severe AR

Determine the viscosity of solutions, Viscosity Viscosity, as you may ...

Viscosity Viscosity, as you may already know, is associated with fluid flow.  It is the internal friction which tends to bring to rest portions of the fluids moving relative to

Define the food and nutrition security, Define the Food and Nutrition Secur...

Define the Food and Nutrition Security? In the previous section, we learnt about the trends in production of food grains in the country and their relationship to population gro

What are the grasslands of north america, What are the grasslands of North ...

What are the grasslands of North America and of South America respectively called? The steppe grasslands of North America are known as prairies. The grasslands of South America

How are inhalation and expiration carried out, Q. How are inhalation and ex...

Q. How are inhalation and expiration carried out? The diaphragm exclusive of mammals and the intercostals muscles can contract or relax varying the volume of the thorax the com

Contagious ecthyma, Contagious ecthyma The contagious ecthyma virus is imm...

Contagious ecthyma The contagious ecthyma virus is immunologically distinct from vaccinia, but similar  to the causative agent of pseudo-cow-pox. It belongs to the genus Parapoxvi

Spinal cord, Spinal cord Spinal cord is a long and cylindrical struc...

Spinal cord Spinal cord is a long and cylindrical structure. It passes through the vertebral column extending all along the dorsal surface of trunk. Vertebrae of the v

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