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

Injection route - administration of drugs, Injection Route An injecti...

Injection Route An injection is an unpleasant experience for a person at any age and is particularly traumatic to the small child who often lives in fear to the next "need

Explain the complications of burns, Explain the Complications of Burns? ...

Explain the Complications of Burns? Most minor burns are superficial and do not cause complications. However, deep second-degree and third-degree burns swell and take more time

How can you explain atp synthesis, Q. How in the respiratory chain do elect...

Q. How in the respiratory chain do electrons from NADH2 and FADH2 passing through cytochromes liberate energy for the ATP synthesis? What is this ATP synthesis called? NADH2and

Cobalt (co) - micronutrients, Cobalt (Co) - Micronutrients The Co conc...

Cobalt (Co) - Micronutrients The Co concentration in the dry matter of plants grown in soil normally lies around 0.02 to 0.5 ppm. In soils the content varies from 1 to 40 ppm.

Patters of cleavage, Patters of Cleavage In most of the animal groups...

Patters of Cleavage In most of the animal groups with spherical or almost spherical egg and little or moderate amount of yolk (micro-or mesolecithal eggs), the first and seco

Explain the interaction of vitamin a with proteins, Explain the interaction...

Explain the interaction of vitamin a with Proteins? Proteins: The protein status of an individual influences vitamin A status and transport because an inadequate protein intake

Explain some features of aspergillus, Explain some Features of Aspergillus?...

Explain some Features of Aspergillus? The identifying features include: 1. Macroscopically Aspergillus colonies are powdery and are of different colours like green, blue, bl

Valves of the heart, Pulmonary valve is surface marked at the sternal end o...

Pulmonary valve is surface marked at the sternal end of the left 3rd costal cartilage. Aortic valve is surface marked at the sternal margin of the left 3rd intercostal space. Mitra

Amoeboid tapetum - tapetum, Amoeboid Tapetum - Tapetum It is also know...

Amoeboid Tapetum - Tapetum It is also known as invasive or periplus modial tapetum. This type of tapetum is more prevalent in the monocotyledons (Arum) than in the dicotyledon

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