A) You are on a workstation in Hawaii. How will you go about discovering a novel
ID: 148070 • Letter: A
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A) You are on a workstation in Hawaii. How will you go about discovering a novel organism with anti-bacterial properties? B) How will you use the difference between eukaryotic and prokaryotic cells to treat bacterial infections? C) How will you exploit the lack of operon to make a novel bacterial protein? D) When and why will you use bacteriostatic versus bactericidal antibiotics? E) How will you exploit the features of a plasmid for making a vaccine or any other product that can be used commercially? F) What are the Mechanisms that lead to anti-bacterial resistance in microbes? G) You are looking at a smear of blood from a dog's ear. How will you go about diagnosing what infection your dog has? H) Connect the functions of bacterial structures to its environment. (Soil, plant, cell.....Explanation / Answer
A) The samples can be collected from different sites, the microbes can be stained by different staining methods and traditional approaches (BIOCHEMICAL, PHYSIOLOGICAL, )can be used for phenotypic characterisation
and molecular methods can be employed as well using the DNA . DNA needs to be isolated, PCR AMPLIFICATION OF 16 S RNA sequence,BLAST against the available database of the known organism. The specific genes involved in detoxification of the pathway needs to be checked by PCR.
Succeptibility assays needs to be performed in presence of different concentrations of any particular drug.
SAMPLE--- PRE-TREATMENT--ISOLATION OF CULTURE--SCREENING FOR MICROBES SHOWING HIGH ANTIMICROBIAL ACTIVITY-- CHARACTERISATION(functional, molecular).
B) Eukayotes and prokaryotes differ in many ways such as the DNA polymerase, RNA polymerase , DNA gyrase, and translational machineries of both differ in terms of structure and function. These features can be utilised to develop potential therapeutics targeting these key regulatory processes.
such as cell wall blockers, RNA pol blockers, Translational inhibitors etc.
D) Bacteriostatic antibiotics can be used when the host resistance is working fine, in that case it will stop the infection without killing the microbe and thus hosts natural defence system will takes care of the infection.
if the infection is too severe and unknown in that case bacteriocidal should be used to completely kill the bacteria.
E) The gene of interest which you want to express coding for the commercial product or vaccine can be cloned into the plasmid along with a strong promoter and other components required for the efficient transcription and translation of the gene. After ligating the gene of interest into the plasmid , the plasmid can be transformed into a competent host cell, and the recombinants will be selected for the presence of the gene of interest by selecting against the antibiotic resistance gene marker. The recombinants selected can be further used for large scale production of specific .target protein which after purification , and characterisation can be tested and marketed commercially.
gene ligation into plasmid---recombinant plasmid transformation into host---- selection of recombinant--- protein expression ---and purification---functional characteristaion--- commercial production on large scale.
F) Mechanisms of antibiotic resistance in microbes is due to many ways--
!) Microbe changes the effectiveness of the drug in many ways
by chemically modifying the drug making it less effective
by blocking the drug entry by increasing membrane thickness
overexpression of transporters to efflux the drug out of the cell
mutating their genetic material, enzymatic machineries etc.
G) Blood smear can be used for detection of presence of antigens in blood by ELISA ,OR Different kits with specified biomarkers can be used as well.
blood smear---staining of the cell types-- microscopic visualisation---for change in cell shape, size and number by counting.
By separating different blood cell types(RBCs, WBCs, Platelets) etc looking at their shape, appearance and their number will tell about any infection.
H) Bacterial structure such as pilli, glycocalyx , cell wall , flagella are present to generate a response against environmental cues. Such as flagella is involved in movement against an attractant or repellant molecule present in environment.
Outer membrane present to resist the harsh outer or inner environmental conditions .
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