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 are the haversian canals, What are the Haversian canals and the Volkma...

What are the Haversian canals and the Volkmann's canals of the bones? Is the osseous tissue vascularized? The Haversian canals are longitudinal canals show in the osseous tissu

Sedimentation, Simple sedimentation: The process of removal of suspende...

Simple sedimentation: The process of removal of suspended, colloidal impurities by allowing water to stand undisturbed in big tanks about 5m deep. Most of the suspended particl

What is catabolism, Q. what is Catabolism? Catabolism is a process of b...

Q. what is Catabolism? Catabolism is a process of breaking down large or complex molecules of food components into smaller components. These supply energy to the cell in a form

Chemical element is the central in the chlorophyll molecule, Q. Which chemi...

Q. Which chemical element is the central in the chlorophyll molecule? The chemical element that is the central in the chlorophyll molecule is magnesium. One atom of magnesium

Describe airway and respiratory support in heart disease, Describe Airway a...

Describe Airway and Respiratory Support in the Newborn ? Like in any other emergency situation a stable airway needs to be established first. Newborns with severe respiratory d

Why is maternal milk important for the immune protection, Why is maternal m...

Why is maternal milk important for the immune protection of the baby? Besides being nutritionally significant, maternal milk participates in the defense of the baby against inf

Human physiology, which bones forms lhe non moving muscle attachment in

which bones forms lhe non moving muscle attachment in

Threshold value of different larval tissues, Threshold Value of Different L...

Threshold Value of Different Larval Tissues Threshold value of dissimilar larval tissues for the thyroid hormone concentration One of the main problems of metamorphosis is

Explain about diabetes mellitus, Explain about Diabetes mellitus Diabe...

Explain about Diabetes mellitus Diabetes mellitus (DM) has relation with all the systems of our body. With some systems it has direct relationship, while with others it has in

Explain brifly ribosomes- endoplasmic reticulum , Explain brifly Ribosomes,...

Explain brifly Ribosomes, Endoplasmic Reticulum, Golgi Apparatus ? Ribosomes, Endoplasmic Reticulum, Golgi Apparatus :  The cytoplasm contains many ribosomes, which are sphe

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