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Enzymes. Enzymes. For this discussion, you will use the virtual laboratory found at: http://glencoe.mcgraw-hill.com/sites/dl/free/0078695104/383930/BL_11.html

Visit the site and read the directions on the left side of the laboratory. Be sure to click on the TV monitor to hear more about enzyme reactions and click on the Information button to read more about enzymes.

You’ll be designing and carrying out an experiment to determine 1) the effect of substrate amount on enzyme activity

Following the directions on the website, you should enter your data into a table (click on the computer monitor and note that you must manually transfer the results from the pop-up window into the table) and then click the graph button. Capture the graph into a file by either using the “Snipping Tool” in your Windows menu, a screenshot of the graph, printing the file to a .pdf or scanning the printed page as a photo.
Post your graph and in your own words explain what conclusions can be drawn from the graph and what impact your results have on biological systems. Be specific and provide real-world examples.

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Enzymes

Plasmid mapping. Plasmid mapping. QUESTION 1

A plasmid was cut with a range of restriction enzymes to produce different sized fragments. These fragments were run out on a gel and the fragment sizes produced are shown below.

Single restriction enzyme digests:

Restriction enzyme	Fragment sizes produced
EcoRI	2670
ScaI	2670
SapI	410, 2260
SspI	740, 940, 990

Double digests Restriction enzymes

Restriction enzymes	Fragment sizes produced
EcoRI and ScaI	2120, 550
EcoRI and SapI	410, 1130
EcoRI and SspI	420, 570, 740, 940
ScaI SapI	410, 580, 1680,

ScaI sspI	130, 740, 810, 990
SapI sspI	180, 230, 560, 710, 990

Triple digest

Restriction enzymes	Fragment sizes produced
EcoRI and ScaI and SapI	410, 550, 580, 1130

Use this information to re-construct a genetic map of the plasmid. Draw this plasmid map clearly for assessment. Be sure to include the exact base positions of all restriction enzymes and label your plasmid. Orientate the plasmid with the EcoRI restriction site at the top of the diagram (at base position 0).

QUESTION 2

Use your plasmid genetic map to answer the following questions

i) What would the shortest distance be between the EcoRI and ScaI restriction sites?

ii) What would the fragment sizes be if you digested the plasmid with EcoRI and SspI and ScaI?

iii) The Lac Z gene begins 150bp away from the EcoRI locus and 700bp from the ScaI locus. What would the distance be from the beginning of LacZ to the nearest SspI locus?

QUESTION 3 Research and explain the structure of the following plasmid features. Explain the reason for their inclusion in a plasmid used in molecular biology applications.

i) M13 locus

ii) T7 promotor

iii) LacZ iv) pUC origin

v) ampicillin resistance ORF

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Plasmid mapping

Tag: enzymes

Enzymes

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