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Molecular genetics 1 win q
The document describes the process of inserting a human insulin gene into bacterial DNA through genetic engineering. First, the insulin gene is cut from human DNA using a restriction enzyme, leaving "sticky ends." A plasmid from bacteria is also cut with the same enzyme. The insulin gene and plasmid are mixed and joined together with the enzyme DNA ligase. The combined DNA is then inserted into E. coli bacteria through applying heat or electric shock to open pores in the bacterial membrane, allowing the plasmid to enter. The engineered bacteria now contains the human insulin gene and can produce human insulin protein.
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Molecular genetics 1 win q
- 1.
- 2. Deoxyribonucleic acid (DNA) •Deoxyribonucleic acid (DNA) is a molecule that carries genetic information. • These genetic information is important for all cellular functions, such as cell division and cell differentiation. • Almost all cells in our body contain DNA inside their nuclei.
- 3. DNA Each DNA molecule consistsof two parallel strands twisted around each other to form a double helix. A molecule of DNA is wrapped around proteins to form a single chromatin thread. During cell division, the chromatin threads coil more tightly to form chromosomes inside the cell nucleus. proteins nuclear pore nucleus nuclear envelope
- 4. What is DNAmade of? Double helix ‘untwisted’ One strand of nucleotides Components of a single nucleotide DNA molecule: a long double helix basedeoxyribose sugar phosphate group Sugar phosphate ‘backbone’
- 5. Basic units ofDNA adenine cytosine guanine thymine deoxyribose sugar phosphate group Bases Nucleotides base joins with the phosphate group and deoxyribose sugar group
- 6. Basic units ofDNA • The basic unit of DNA is a nucleotide. • Each nucleotide is made of - a sugar called deoxyribose; - a phosphate group; and - a nitrogen-containing base, all joined together • The four bases of nitrogen-containing bases are - adenine (A) - thymine (T) - cytosine (C) adenine cytosine guanine thymine deoxyribose sugar phosphate group Bases Nucleotides adenine nucleotide thymine nucleotide guanine nucleotide cytosine nucleotide base joins with the phosphate and deoxyribose sugar group
- 7. The building blocksof DNA • Nucleotides are joined together to form long chains called polynucleotides. • Each gene is made up of a sequence of nucleotides. This sequence can vary. bases sugar-phosphate backbone polynucleotide
- 8. Rule of basepairing • The bases of one strand form bonds with bases of the other strand according to the rule of base pairing. • Adenine (A) bonds with thymine (T), while cytosine (C) always bonds with guanine (G). • Bases that bond with each other are known as complementary base pairs. base pair
- 10. Guide to bea God! 1) Colour each of the individual structures on the worksheet with a different colour: Example: adenine = red thymine = green guanine = blue cytosine = yellow phosphate = brown deoxyribose = purple
- 11. Guide to bea God! 2) Cut out each structure. 3) Using the small symbols (squares, circles and stars) on the structures as guides, line up the bases, phosphates and sugars. 4) Glue the appropriate pairs together to form nucleotides.
- 12.
- 13. Guide to bea God! 5) Construct the right side of your DNA molecule by putting together in sequence a cytosine, thymine, guanine and adenine nucleotide. 6) Complete the left side of the DNA ladder by adding complementary nucleotides or nucleotides that fit. Your finished model should resemble a ladder.
- 14. Guide to bea God! 7) To show replication of your model, separate the left side from the right side on your desk, leaving a space of about 15 to 20 cm. 8) Using the remaining nucleotides, add to the left side of the model to build a new DNA molecule. Do the same with the separated right side.
- 15. Questions?!?! 1) When constructingthe DNA molecule, what did you notice about the orientation of the two strands? 2) What DNA strand would bond opposite? 3) What is a similarity and a difference in DNA between Homo sapien and Blattaria?
- 16. The DNA doublehelix The DNA molecule has a spiral structure known as the double helix. Both strands of DNA that run in opposite directions are twisted to form this double helix. a base pair sugar-phosphate backbone coiling of DNA double helix structure of DNA
- 17. Genes • A DNAmolecule contains many genes along its length. • A gene is a small segment of DNA which controls the formation of a protein, such as an enzyme. gene DNA molecule
- 18. Genes • Each genestores a message that determines how a protein should be made in a cell. • The message stored by a gene is known as the genetic code. • Proteins are responsible for the development of certain characteristics in the body. gene DNA molecule part of a DNA molecule unzipped to show a gene M E S S A G E a gene is a segment of DNA protein coded by the gene
- 19. Structure of agene • Each gene consists of two polynucleotide chains. One of the chains determines the type of protein made. This chain is called the template. • The template is made up of a sequence of nucleotide bases. • Three sequential bases code for one amino acid. This is known as the triplet code or codon. DNA template Process of decoding and protein synthesis polypeptide made of five amino acids triplet code/ codon
- 20. How are proteinsmade? • Proteins in the cell are made through a two-step process — transcription and translation. • Transcription occurs when the message in the template has to be copied into an RNA molecule called messenger RNA (mRNA). • Transcription occurs in the nucleus. • Three bases in the mRNA made up a codon. DNA template TranscriptionmRNA - RNA contains U (uracil) instead of T (thymine) codon
- 21. How are proteinsmade? • The mRNA moves out of the nucleus and carries the message to the cytoplasm. • A ribosome helps to translate the sequence of codons on the mRNA into a protein molecule. mRNA polypeptide Translation
- 22. Comparing DNA andRNA DNA (double helix) RNA Sugar unit is deoxyribose. Sugar unit is ribose. Nitrogen-containing bases are adenine (A), thymine (T), cytosine (C) and guanine (G). Nitrogen-containing bases are adenine (A), uracil (U), cytosine (C) and guanine (G). Permanent molecule in the nucleus Temporary molecule that is made when needed Found only in nucleus Found in nucleus and cytoplasm
- 23. 1 part ofa gene Transcription and Translation
- 24. First, the geneunzips. 1 part of a gene Transcription and Translation
- 25. template mRNA molecule is made Oneof the strands in the gene is used as the template to make mRNA. This is transcription. The mRNA molecule copies the genetic code in the DNA template, following the rule of base pairing. 1 Note that mRNA does not contain T (thymine). A (adenine) in DNA pairs with U (uracil) in mRNA. Transcription and Translation
- 26. mRNA molecule is made ribosome mRNA nuclear envelope ThemRNA leaves the nucleus and attaches to a ribosome in the cytoplasm. 2 nuclear pore Transcription and Translation
- 27. tRNA • Inthe cytoplasm are amino acids and transfer RNA (tRNA). Transfer RNA or tRNA is another RNA molecule also needed for protein synthesis. • tRNA molecules have amino acids attached to one end of their structure.Each tRNA is very specific and attaches only to its own amino acid For example, a tRNA with the anticodon UAC always attaches to the amino acid M. • Each tRNA also has three bases at one end. This is an anticodon that can bind to complementary codons on mRNA. cytoplasm amino acids 3 anticodon Transcription and Translation
- 28. codon The anticodons ontRNA bind with their respective codons on mRNA. tRNA amino acid attached to tRNA peptide bond ribosome 4 • Translation starts with mRNA attaching to a ribosome. • The first two tRNAs together with their amino acids also fit into the ribosome. They attach to the codons on the mRNA according to the rule of base pairing. • A peptide bond is formed between the two amino acids. Transcription and Translation
- 29. 5 peptide bond between aminoacids first tRNA is released a new tRNA fits into the ribosome • Once the peptide bond is formed between the first two amino acids, the ribosome moves along one codon to the right of the mRNA. • As the ribosome moves to this position, the first tRNA is released. • At the same time, the third tRNA and its amino acid slots into the ribosome. codon ribosome moves along the mRNA strand Transcription and Translation
- 30. 6 another amino acid isattached to the chain direction of movement of ribosome • Another amino acid is attached to the chain. Transcription and Translation
- 31. polypeptide formed • Theprocess continues as the ribosome moves along the mRNA. • At the end of the mRNA is a stop codon such as UGA, UAA or UAG. A stop codon does not have any tRNA with complementary codons. This means that anticodons ACU, AUU or AUC do not exist. • Eventually, the whole chain of polypeptide is produced. The ribosome leaves the mRNA. 7 Transcription and Translation
- 32. Transcription and Translation polypeptideformed • The ribosome may attach to the same mRNA for another round of translation. 8
- 33. insulin gene • Obtainthe human chromosome containing the insulin gene. 1 How the human insulin gene is inserted into bacterial DNA Genetic Engineering
- 34. insulin gene • Obtainthe human chromosome containing the insulin gene. • Cut the gene using a restriction enzyme. This enzyme cuts the two ends of the gene to produce ‘sticky ends’. 1 cut by restriction enzyme How the human insulin gene is inserted into bacterial DNA fragment of DNA containing the insulin gene sticky end Genetic Engineering
- 35. insulin gene • Obtainthe human chromosome containing the insulin gene. • Cut the gene using a restriction enzyme. This enzyme cuts the two ends of the gene to produce ‘sticky ends’. • Each ‘sticky end’ is a single strand sequence of DNA bases. These bases can pair with complementary bases to form a double strand. 1 cut by restriction enzyme fragment of DNA containing the insulin gene sticky end How the human insulin gene is inserted into bacterial DNA Genetic Engineering
- 36. insulin gene • Obtaina plasmid from a bacterium. 2 cut by restriction enzyme fragment of DNA containing the insulin gene sticky end plasmid How the human insulin gene is inserted into bacterial DNA Genetic Engineering
- 37. insulin gene • Obtaina plasmid from a bacterium. • Cut the plasmid with the same restriction enzyme. This produces complementary sticky ends. 2 cut by restriction enzyme fragment of DNA containing the insulin gene sticky end plasmid cut by same restriction enzyme sticky ends How the human insulin gene is inserted into bacterial DNA Genetic Engineering
- 38. insulin gene • Mixthe plasmid with the DNA fragment containing the insulin gene. 3 cut by restriction enzyme fragment of DNA containing the insulin gene sticky end plasmid cut by same restriction enzyme sticky ends How the human insulin gene is inserted into bacterial DNA Genetic Engineering
- 39. insulin gene • Mixthe plasmid with the DNA fragment containing the insulin gene. • Add the enzyme DNA ligase to join the insulin gene to the plasmid. 3 cut by restriction enzyme fragment of DNA containing the insulin gene sticky end plasmid cut by same restriction enzyme sticky ends insulin gene inserted into plasmid How the human insulin gene is inserted into bacterial DNA DNA ligase Genetic Engineering
- 40. insulin gene • Mixthe plasmid with E. coli bacteria. 4 cut by restriction enzyme fragment of DNA containing the insulin gene sticky end plasmid cut by same restriction enzyme sticky ends insulin gene inserted into plasmid E. coli bacterial DNA How the human insulin gene is inserted into bacterial DNA DNA ligase Genetic Engineering
- 41. Genetic Engineering insulin gene •Mix the plasmid with E. coli bacteria. • Apply temporary heat or electric shock. This opens up pores in the cell surface membrane of each bacterium for the plasmid to enter. 4 cut by restriction enzyme fragment of DNA containing the insulin gene sticky end plasmid cut by same restriction enzyme sticky ends insulin gene inserted into plasmid plasmid bacterial DNA plasmid enters the bacterium trangenic bacterium E. coli bacterial DNA How the human insulin gene is inserted into bacterial DNA DNA ligase