BIOL1181 2250 Assignment 1: Applied Theory (WEEKS 1-3) ● Please read the instructions that have been posted on CANVAS before completing this assessment. Please use either Calibri (Font Size 11) or

Arial Font (Size 10) to type your answers. Adhere to the space provided for each question. Question 1. You have recently joined a biotech company that specialises in the cloning and expression of human hormones for therapeutic use [Total = 12 marks]. Your first task it to express the Human Growth Hormone, whose structure is depicted below: Genomic DNA 5 5' size (bp) cDNA GH protein BIOL1181 Assignment 1: Applied Theory (WEEKS 1-3) Total Marks = 35 nukleotides 5175 5234 I exons Answer: ▼ IVS 1 Answer: 10 261 5'NT 5495 | ✓ 161 5655 5865 5984 6077 | ✓ ✓ IVS 2 ✓ IVS 3 209 120 93 31 40 55 IV 6241 165 • 65 IVS 4 253 6495 1 ▼ -3-23 1-31 32-71 72-126 127-191 (aa) V 195 6689 (21) You need to provide your supervisor with a strategy to clone this gene in an expression vector for expression in a bacterial cell system. You need to: 3'NT Poly A a. Explain to your supervisor which starting material you will use (genomic DNA? mRNA?) to prepare your insert, and why? [2 marks]. b. List the elements that must be present in this vector for the successful expression of the bacterial system, and why these elements are required? [4 marks]. BIOL1181 2250 Assignment 1: Applied Theory (WEEKS 1-3) c. List the mains experimental steps required for cloning your insert of interest into the vector, starting from extraction of the relevant nucleic acid from cells (as you have defined under Question 1a) and finishing with a pure population of bacteria that has acquired the plasmid. Indicate which enzyme(s) is(are) are required at each step [6 marks]? You can use a schematic. Answer: