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

nutrient stability as affected by processing of feeds, Nutrient stability ...

Nutrient stability as affected by processing of feeds Nutrient stability under normal conditions of storage and processing is one of the important factors determining the effe

Explanation of glomerular filtration, Question 1) Mentioning the 3 diff...

Question 1) Mentioning the 3 different processes in the formation of urine Explanation of Glomerular filtration Explanation of Tubular reabsorption Explanation of T

Explain the expanding frontiers of nutrition, Explain the Expanding Frontie...

Explain the Expanding Frontiers of Nutrition? While the essential nutrients mentioned in Table are indispensable and our diets need to provide them all in adequate quantities,

How do grasslands differ from savannas, How do grasslands differ from Savan...

How do grasslands differ from Savannas? Name the two main categories of plant forms that dominate the desert vegetation.

What is pinocytosis, Pinocytosis: - The  ingestion of dissolved materials  ...

Pinocytosis: - The  ingestion of dissolved materials  by  endocytosis.  The cytoplasmic membrane invaginates and pinches off placing small  droplets of fluid  in  a  pinocytic vesi

Define the chromosome region, Q. How is the chromosome region where the cen...

Q. How is the chromosome region where the centromere is located called, and how chromosomes classified in the relation to the position of their centromere? The chromosome regio

Physiological adaptaion, what are the physiological adaptation shown my pre...

what are the physiological adaptation shown my prey as defence

Planning the nursing care for infective endocarditis, Planning the Nursing ...

Planning the Nursing Care   Provide bed rest  Administer antibiotics as advised Prevent infection  Implementation of Nursing Care  Provide Bed Rest   Sin

Explain the nutritional and functional role of magnesium, Minerals :- Magne...

Minerals :- Magnesium    Food Source      Whole grains, nuts, legumes, green leafy vegetables Nutritional Functional role Essential nutrient: Deficiency is rare Co

Explain the distant osteogenesis, Explain the Distant Osteogenesis In t...

Explain the Distant Osteogenesis In this an extracellular matrix establishes the implant surface contact. The osteogenic cells line the host bone surface. The blood supply to t

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