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

Indicator organism, a) What are the needsfor an organism to be an indicator...

a) What are the needsfor an organism to be an indicator organism? b)  In Australia, National Parks and coastal beaches attract various visitors during holiday periods.  EPA is

What do you mean by bone implant interface, Q. What do you mean by Bone Imp...

Q. What do you mean by Bone Implant Interface? It consists of remodelled bony tissue. For making it strong implant should not be overloaded during its organization period i.e.

Iron deficiency anaemia, Iron Deficiency Anaemia   This is the most com...

Iron Deficiency Anaemia   This is the most common nutritional and haematologic disorder  in infancy and chhildhood in developing countries. It is caused by lack of sufficient i

What is biochemical characteristics, What is Biochemical Characteristics? ...

What is Biochemical Characteristics? In the last section we have learnt about the importance of biochemical tests. It must be clear now that biochemical tests are used to -

Caryopsis - development of fruit, Caryopsis - Development of Fruit In ...

Caryopsis - Development of Fruit In cereals each carpel has one ovule and therefore the mature fruit has, just one seed. During maturation, very little or no cell divisions ar

What is the dental implants option, Dental implants option It is import...

Dental implants option It is important that the type of prosthetic options selected for the patient is not only cost effective, but also must  be predictable and restores the e

What do we mean by culture media, What Do We Mean By Culture Media? Cul...

What Do We Mean By Culture Media? Culture media, you would realize, is a solid or liquid preparation containing all the nutrients required by microbes for growth. So then, what

Breeder select for in a cereal crop, What characteristics might a breeder s...

What characteristics might a breeder select for in (i) a cereal crop, (ii) a farm animal? (i) In a cereal variety, a breeder might select for high yield; disease-, frost- or dr

Explain phytoestrogens, Phytoestrogens:- A group of compounds considere...

Phytoestrogens:- A group of compounds considered to offer protection against cancer.

The fragility of erythrocytes, The fragility of erythrocytes The fragil...

The fragility of erythrocytes The fragility of erythrocytes is impaired in the absence of NADPH generation due to the deficiency of glucose-6-phosphate dehydrogenase thereby ca

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