Transgenic Fly Biology Lab

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View This

Promega video about reporter genes assays:

https://www.youtube.com/watch?v=xqTudg1WdmY

Youtube video about how to make transgenic flies:

https://www.youtube.com/watch?v=1CJfj1GG_f8

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Transgenic organisms are defined as a genetically modified organisms that contains DNA from two different genomes. A transgenic organism (or knock-in organism) contains a gene which is overexpressed for the purpose of ascertaining the gene function and location.  The method of creating a transgenic organism is the same as creating a knock-out organism.  A plasmid vector must be used for inserting the gene of interest into the genome.  The vector is introduced into the gametic cells of the organism and the experssion is confirmed in the offspring.

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“Make Transgenic Flies” will be completed first then do the “Use Transgenic Flies”.

Copy/past this link into your browser. Click on “Launch Interactive.” Complete the lab and answer the questions on the worksheet. Remember to read all the information.

https://www.biointeractive.org/classroom-resources/transgenic-fly-virtual-lab

Make Transgenic fly lab 1. (1 pt) You must submit a screen shot of your successful mutant flies at the end of the “Make Transgenic fly lab”. This screenshot will be submitted to a Discussion after completing the lab. When submitting the worksheet – please answer: Where you successful in your experiment? What went wrong? 2. (0.5 pts) What gene are you attaching to the period gene to visualize when transcription occurs? Which organism does this gene come from? 3. (0.5 pts) What key molecular “tool” is used to create the construct of per/luc genes that will jump into the Drosophila genome? 4. (0.5 pts) How will we know if the construct has incorporated into the Drosophila genome? 5. (1 pt) Complete part A of Performing a sequence analysis in the “Make Transgeneic lab” to confirm that the correct period gene is incorporated into the plasmid construct. In the BLAST sequence, what color means a good match? What chromosome is period gene found in Drosophila? Testing Transgenic flies 1. (0.5 pts) About what time of the day does the fly have peak luciferase expression? 2. (0.5 pts) Does the Per gene continue to oscillate regardless of external light/dark cues? 3. (0.5 pts) What time did the peak luciferase expression become after the shift in light/dark cycle?

 
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Discoveries in the War on Cancer

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BIOL 101

Individual Assignment 3: 10 Discoveries in the War on Cancer

1. Scientists have always dissected lymph nodes nearby a cancerous tumor to see if the cancer has begun to spread to other areas of the body. The most obvious nearby lymph node is chosen. Scientists have discovered that in women with breast cancers that are 5 cm in diameter or smaller, nothing is gained by additional lymph node dissections in the axillary region (armpit) of the body. Dissecting the node nearest the tumor provides adequate information by itself.

2. Malignant brain tumors in adults are fast-growing cancers with median survival rates of 15 months, even with aggressive treatment. Researchers have been searching for genetic “signatures” (characteristic groups of cancer-causing genes) that could help in defining the kind of brain tumor the patient has. They hope to be better able to predict the course of the disease and the patient’s response to treatment.

3. Molecular geneticists have surveyed the entire human gene collection (the “genome”) in patients with neuroblastoma , a childhood cancer that forms in the adrenal glands and nerve tissue. Results of a genome-wide association study indicate that a specific region on human chromosome 1 is associated with the development of this cancer. This chromosome region contains a section of DNA that can vary in copy number from person to person, and neuroblastoma patients were more likely than control subjects to have fewer than normal copies of this DNA.

4. Tobacco smoking is the leading cause of preventable deaths worldwide. It is a risk factor for lung cancer and several other types of cancer. Results of analysis of the entire human gene collection (the “genome”) support some previous findings that a region of human chromosome number 15 contains one or more genes that are associated with smoking intensity (the number of cigarettes smoked per day) and the closely related trait of nicotine dependency.

5. Molecular biologists have developed a new sequence of human genes called an ankyrin insulator sequence. You place a new corrected or therapeutic gene within this sequence. Its role is to create an active area on a human chromosome where the new gene can work efficiently no matter what chromosome it lands on.

6. Molecular biologists have taken nanoparticle -sized spheres and used them to deliver a cell-killing toxin from bee venom to tumors in mice, substantially reducing tumor growth without harming normal body tissues. Nanoparticles are known to concentrate in solid tumors because blood vessels in tumors show “enhanced permeability and retention effect” or EPR. Hence, substances such as nanoparticles escape more readily from the bloodstream into tumors, and the generally poor drainage of lymph from tumors further helps trap the particles in tumor tissue.

7. Organic chemists are exploring structural variations of the organic compound avobenzone (1-(4-Methoxyphenyl)-3-(4-tert-butylphenyl) propane-1,3-dione) for inclusion in sunblock products. Avobenzone is known for its ability to absorb a broad spectrum of ultra-violet radiations including UVb light (known to enhance the frequency of basal cell and squamous cell carcinomas (skin cancers)) and UVa rays thought to increase the frequency of melanoma cancers. New variations in the structure of avobenzone are hoped to retain the ability to absorb harmful UV radiation while having an increased stability in the presence of that radiation.

8. Immunologists are investigating ways to destroy lymphocytes (white blood cells of the immune system) that have become cancerous (lymphomas). A current drug Rituxamab contains antibodies that bind to the surfaces of these lymphocytes, setting them up for destruction by the cancer patient’s own immune system. They are currently seeking ways to modify the antibody’s structure so that it will attract the cancer patient’s “natural killer” (NK) cells to the lymphocytes. Success of this project will bring a multi-faceted immune response against lymphomas and hasten its destruction.

9. Biochemists are analyzing the many, many components of red meat (beef, pork) to determine which component, if any, will cause increased colorectal cancer rates in mice when the component is administered orally. Studies have shown that higher colorectal cancer rates in humans are associated with higher consumption rates of red meat.

10. Virologists are modifying lentiviruses as vectors for carrying proto-oncogenes into cancer-transformed cells in culture. They are developing this virus for inserting the ras proto-oncogene directly into its correct location in the genome. The correct ras gene will already be linked to human DNA to either side of it and complexed with a recombination enzyme that will insert it into its correct location within the human genome. At the same time, the recombination enzyme will excise the defective oncogenic form of ras. The cells in culture should again come under normal hormonal control and require extra-cellular signals in order to continue dividing.

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Skeletal System

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1. The skeleton is divided into 2 sections. The __________________ skeleton which includes the skull, ribs, sternum, and vertebrae, and the _______________________ skeleton which is mainly the limbs.

2. Go to Busy Bones – Virtual Bones Lab then click on the “Skeleton Viewer” to investigate the names and positions of the various bones. Label the skeleton below.

3. How many ribs make up the rib cage?

4. What is the name of the bone labeled “#2”?

5. What two major organs does the rib cage protect?

6. If you were punched in the stomach, would your ribs protect you? Explain!

7. How many vertebrae can you count in this spine?

8. Label the 7 cervical vertebrae on your diagram.

(C1, C2, C3, C4, C5, C6, C7)

9. Label the 12 thoracic vertebrae on your diagram.

(T1, T2, T3, T4, T5, T6, T7, T8, T9, T10, T11, T12)

10. Label the 5 lumbar vertebrae on your diagram.(L1, L2, L3, L4, L5)

11. Label the sacrum and coccyx on your diagram.

12. How many cartilage disks are in your spine?(Hint: you will need to count)

13. Identify the types of joints:

a. Elbow =

b. Skull =

c. Vertebrae =

d. Hip =

14. Go to Busy Bones – Virtual Bones Lab then click on the “Virtual Bone Microscope”

15. Click on slide 1

a. What is the other name for Trabecular bone?

b. Describe the characteristics of

i. Spongy bone

ii. Compact bone

16. Click on slide 2

a. Describe the epiphysis and its function.

b. Describe the growth plate and its function.

17. Click on slides 3 and 4

a. What do the osteoblasts do?

18. Click on slide 5, 7, and 8

a. Describe what is in the Haversian canals.

b. Describe the canaliculi and their function.

19. Click on slide 6

a. Describe the trabeculae and what fills this.

1. The skeleton is divided into 2 sections. The __________________ skeleton which includes the skull, ribs, sternum, and vertebrae, and the _______________________ skeleton which is mainly the limbs.

2. Go to Busy Bones – Virtual Bones Lab then click on the “Skeleton Viewer” to investigate the names and positions of the various bones. Label the skeleton below.

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Posted: A Day Ago
Due: 12/10/2019
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2420-Lab 7- Microbial Growth

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2420-Lab 7- Microbial Growth: Temperature, Oxygen and Osmotic Balance

Directions:

Answer following questions after reading the information and watching the video from the link below. Use color RED or BLUE for your answers. Submit the completed document on eCampus for grading. Refer to

· the textbook chapter 4 (sections 4.1, 4.3 and 4.4) (Nester- McGraw Hill)

· Lab Manual by Dr. Su, pages (optional)

· Link: Environmental Influences of Bacterial Growth, Virtual Edge Experiment- 5A-B

· pH Requirement of bacteria

Bacterial Growth:

Read the information from textbook from chapter 4 review sections 4.1, 4.3 and 4.4 to answer the following questions.

Watch the following video:

· For background: Bacterial Growth

1. Label the phases of growth on the following curve:

2. What happens to bacteria in the phases labeled as A, B, C and D in the above figure?

A:

B:

C:

D:

3. If we add more nutrients and space at the end of the exponential phase for a growing bacterial culture, what will happen?

Oxygen Requirement of bacteria

Read the information from textbook from chapter 4 review sections 4.3-4.4 to answer the following questions.

Watch the following videos:

· For background: Oxygen Requirement of bacteria

· For experiment: Oxygen requirement of bacteria using soft agar

Results of the bacterial oxygen requirement of bacteria:

4. Why can only some organisms tolerate oxygen?

 

5. Describe the kind of bacteria shown in the figure above in terms of oxygen (concentration) requirement and special enzymes.

 

6. Which oxygen related enzymes are present or absent in the bacteria from tubes A-E?

 

Temperature Requirements of Bacteria:

Read the information from textbook from chapter 4 review sections 4.3-4.4 to answer the following questions.

Watch the following videos:

· For background: Temperature Requirement of bacteria

· For experiment: Temperature Influence on bacterial growth

Temperature tolerance of bacteria

Write the correct term for bacteria growing under the following conditions

7. Most medically important bacteria

8. Bacteria growing on or inside your body

9. Bacteria growing in your refrigerator

10. Bacteria growing in the arctic ocean in winter

11. Bacteria growing in hot springs in Arkansas

12. Bacteria growing in geysers

13. Why are bacteria not able to grow at temperature higher than the maximum tolerance limit?

14. Do bacteria grow at the temperatures lower than the minimum tolerance limit? Why or why not? Explain

Osmotic Balance Requirements of Bacteria:

Read the information from textbook from chapter 4 review sections 4.3-4.4 to answer the following questions.

Read the following:

· For experiment: Osmotic Influences

Osmotic tolerance of bacteria

Halophiles : On human body, Staphylococcus aureus is known to tolerate sweat. These bacteria may cause skin infections if athletic gear is shared among athletes without proper sanitization. In nature we will find bacteria, algae and fungi in water bodies or soil with higher salt concentrations. These organisms come under a group called “halophiles”.

Habitat : Halophiles are usually found in salt lakes, salt marshes, subterranean salt deposits, dry soil, salted meat and hypersaline seas.

3 groups based on the salt concentration tolerance :

· Halophiles-2-5% salt required for growth

· Moderate halophiles-5-20% salt required for growth

· Extreme halophiles-20-30% salt required for growth

Write the correct term for bacteria growing under the following conditions

15. Based in your knowledge about tonicity, for cells to survive they have to be surrounded by ______tonic environment.

 

16. Why would cells not survive in hypotonic or hypertonic environment?

2420-Lab 7- Microbial Growth: Temperature, Oxygen and Osmotic Balance

Directions:

Answer following questions after reading the information and watching the video from the link below. Use color RED or BLUE for your answers. Submit the completed document on eCampus for grading. Refer to

· the textbook chapter 4 (sections 4.1, 4.3 and 4.4) (Nester- McGraw Hill)

· Lab Manual by Dr. Su, pages (optional)

· Link: Environmental Influences of Bacterial Growth, Virtual Edge Experiment- 5A-B

· pH Requirement of bacteria

Bacterial Growth:

Read the information from textbook from chapter 4 review sections 4.1, 4.3 and 4.4 to answer the following questions.

Watch the following video:

· For background: Bacterial Growth

1. Label the phases of growth on the following curve:

2. What happens to bacteria in the phases labeled as A, B, C and D in the above figure?

A:

B:

C:

D:

3. If we add more nutrients and space at the end of the exponential phase for a growing bacterial culture, what will happen?

Oxygen Requirement of bacteria

Read the information from textbook from chapter 4 review sections 4.3-4.4 to answer the following questions.

Watch the following videos:

· For background: Oxygen Requirement of bacteria

· For experiment: Oxygen requirement of bacteria using soft agar

Results of the bacterial oxygen requirement of bacteria:

4. Why can only some organisms tolerate oxygen?

 

5. Describe the kind of bacteria shown in the figure above in terms of oxygen (concentration) requirement and special enzymes.

 

6. Which oxygen related enzymes are present or absent in the bacteria from tubes A-E?

 

Temperature Requirements of Bacteria:

Read the information from textbook from chapter 4 review sections 4.3-4.4 to answer the following questions.

Watch the following videos:

· For background: Temperature Requirement of bacteria

· For experiment: Temperature Influence on bacterial growth

Temperature tolerance of bacteria

Write the correct term for bacteria growing under the following conditions

7. Most medically important bacteria

8. Bacteria growing on or inside your body

9. Bacteria growing in your refrigerator

10. Bacteria growing in the arctic ocean in winter

11. Bacteria growing in hot springs in Arkansas

12. Bacteria growing in geysers

13. Why are bacteria not able to grow at temperature higher than the maximum tolerance limit?

14. Do bacteria grow at the temperatures lower than the minimum tolerance limit? Why or why not? Explain

Osmotic Balance Requirements of Bacteria:

Read the information from textbook from chapter 4 review sections 4.3-4.4 to answer the following questions.

Read the following:

· For experiment: Osmotic Influences

Osmotic tolerance of bacteria

Halophiles : On human body, Staphylococcus aureus is known to tolerate sweat. These bacteria may cause skin infections if athletic gear is shared among athletes without proper sanitization. In nature we will find bacteria, algae and fungi in water bodies or soil with higher salt concentrations. These organisms come under a group called “halophiles”.

Habitat : Halophiles are usually found in salt lakes, salt marshes, subterranean salt deposits, dry soil, salted meat and hypersaline seas.

3 groups based on the salt concentration tolerance :

· Halophiles-2-5% salt required for growth

· Moderate halophiles-5-20% salt required for growth

· Extreme halophiles-20-30% salt required for growth

Write the correct term for bacteria growing under the following conditions

15. Based in your knowledge about tonicity, for cells to survive they have to be surrounded by ______tonic environment.

 

16. Why would cells not survive in hypotonic or hypertonic environment?

 
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