Please answer parts d and e **Image Transcription for Educational Website****Diagram Overview:**The diagram is a circular map of the plasmid pUC19, which is 2,686 base pairs (bp) in length. The map highlights various restriction enzyme cut sites along the plasmid, including:- BamHI (2683 bp)- Eco01091 (2674 bp)- AatII - ZraI (2617 bp)- BsaW1 (2542 bp)- SspI (2501 bp)- AclI (2297 bp)- XmaI (2294 bp)- BglI (2215 bp)- ScaI (2217 bp)- PvuI (2066 bp)- AseI (2016 bp)- EcoRV (1953 bp)- MscI (1924 bp)- PmlI (1919 bp)- AvaII (1822 bp)- NruI (1830 bp)- BspHI (1774 bp)- BsrFI (1779 bp)- BbvCI (1753 bp)- MluI (1744 bp)- AhdI (1694 bp)- BceAI (1292 bp)- AlwNI (1217 bp)- BbvCI (1105 bp)- BlpI (1015 bp)- AflIII - PciI (806 bp)- BspDI/SapI (683 bp)- TfiI (671 bp)- PsiI (628 bp)- DpnII (403 bp)Additionally, major structural features are labelled:- **lacZ**: The LacZ alpha-peptide region- **Ampr (Ampicillin resistance gene)**: Confers resistance to ampicillin- **ori (Origin of replication)**: Required for plasmid replicationThe diagram lists an MCS (Multiple Cloning Site) near the lacZ region, with various enzymes like ApaI, KpnI, and SmaI both on the map and highlighted in a callout box.**Text Content:**a) The plasmid codes for three elements, lacZ, Ap^R, and ori. What are these three things?b) Which restriction enzyme cut sites would you use to clone your blue pigment gene into pUC19 and why?c) You digest the plasmid and your blue pigment gene with the restriction enzymes

The insertion of a gene into a foreign gene is known as gene cloning or genetic engineering. Prokaryotes also have plasmids, which are extrachromosomal DNA that can be genetically modified. The plasmids that are readily accessible in the marketplace have restriction sites that allow for the insertion of foreign DNA following nicking by an appropriate restriction enzyme. The plasmid contains a selectable marker gene, such as an antibiotic resistance gene, that can be used to identify cells that have undergone genetic modification.d. The selectable markers are used to know which host cells have taken the plasmid. The selectable marker is usually an antibiotic resistance gene which is absent in the host. Hence the host without plasmid can not grow on antibiotic-containing plates. When host cells take up the plasmid, the transformed cells gain antibiotic resistance and can grow on antibiotic-containing plates. Hence we use antibiotic-containing plates to select transformants from the non-transformants. All the cells which have taken the plasmid can grow on antibiotic-containing plates irrespective of whether the insert is present in the plasmid or not. Here, the selectable marker gene is the AmpR gene which codes for the ampicillin resistance gene. Hence all the cells which grow on ampicillin antibiotic-containing plates have taken the plasmid. e. Blue-white screening is used to identify the positive and negative clones by adding X-gal to the growth medium. The positive clones form white colonies as the insert has been inserted in the lacZ gene, hence it can not produce the X-gal metabolizing enzyme. We insert the gene of interest into MCS so that it disrupts the Beta-galactosidase gene, which does not produce the whole-length enzyme if there is an insert, interrupting the gene sequence. When the lacZ gene is intact, in negative clones (without insert), Beta-galactosidase is produced, and it will cleave X-gal to produce a chromogenic material, resulting in a blue-coloured compound, suggesting that blue colonies are negative clones where the gene has not been introduced in the lacZ gene.