Introductory to Molecullar Biology

Rizki Meizikri Wahyu Tri Novriansyah Muhammad Farid Shofi Faiza Deby Nelsya Eka Putri Muhammad Ihsan Fachruddin Akbara Pradana Ghucyka Jhonelta Afrilizia Putri Carlven Lenim

Group 10

(1010311010) (1010312013) (1010312041) (1010312069) (1010312097) (1010313013) (1010313047) (1010313082) (1010313095) (1010314010)

Riri, 18, read news on the mass media about a terrorist shot to death by the police. To check the validity of the terrorist·s identity, a DNA test should be done. Riri who is currently learning about molecullar biology is very excited with the news. She recalls her highschool lesson about DNA·s function which can maintain its characteristic through replication process, and mutation may occur if DNA is exposed to radiation. Riri learns from several sources that DNA transcription produces RNA that will be translated in protein synthesis. DNA recombination, a genetical engineering technique, can also help producing stem cell, vaccines, medicines, and even cloning which becomes ethical dilemma. How do you describe these problems?

Biomolekuler rRNA Cloning Stem Cell

Rekayasa Genetika



Recombinant DNA

Vaccine DNA mRNA tRNA






1. Students Able to Explain About Structure and Function of DNA and RNA
Nitrogen Base


Phosphate Group



Thymine (T) Pyrimidines

Cytosine (C)

Adenine (A) Purines

Guanine (G)

Nitrogenous base (A, G, C, or U) Phosphate group


Uracil (U)

Sugar (ribose)

Differences Between DNA and RNA
Differences Shape Sugar Pyrimidines Function(s) Stability Location DNA Double Helix Deoxyribose Cytosine, Thymine Protein synthesis, Inheritance Stabile Nucleus RNA Single Strand Ribose Cytosine, Uracil Protein synthesis Less stabile Nucleus and Cytoplasm

2. Students Able to Understand Factors That Affect DNA Stability
y Hydrogen Bonding y Thermal Melting y Phosphate bonding y pH y Radiation, ex : UV, infra red, X-Ray y Virus DNA and RNA

3. Students Able to Explain DNA Replication Process

DNA polymerase molecule Parental DNA Daughter strand synthesized continuously Daughter strand synthesized in pieces

How Nucleotides Added in DNA Replication

Leading Strand
y Primase enzyme put a primer RNA on the strand y DNA polymerase III begin adding nucleotides y DNA polymerase I transforms the primer RNA to DNA y The replication continues until the end of the strand

Lagging Strand
Primase enzyme put a primer RNA on the strand DNA polymerase III begin adding nucleotides Primase enzyme put another primer RNA on the strand DNA polymerase III stop its activity and move to the new primer RNA ‡ DNA polymerase III stops when it meets the first primer RNA, leaving the strand unfilled. The blank space is called Okazaki Fragment ‡ Primer RNA will be transformed to DNA by DNA polymerase I, so that ligase can bind it with the DNA made by DNA polymerase III ‡ ‡ ‡ ‡

4. Students Able to Explain Protein Synthesis Process
Transcribed strand DNA



Start codon


Stop codon


RNA polymerase DNA of gene DNA Promoter DNA Initiation Terminator DNA Cap RNA transcript with cap and tail Transcription Addition of cap and tail Exon Intron Exon Intron Exon

Introns removed


Elongation mRNA

Exons spliced together

Coding sequence NUCLEUS Termination Growing RNA CYTOPLASM Completed RNA RNA polymerase

5. Students Able to Explain DNA Mutation
Types of DNA mutation: y Missense Mutation Nitrogen base changed, amino acid changed (ex: Sickle Cell Anemia) y Nonsense Mutation Nitrogen base changed, codon turned into stop codon y Silent Mutation Nitrogen base changed, amino acid doesn·t change
y Frameshift Mutation

Abnormal number of nitrogen base y Splice Site Mutation Failure in intron removal

6. Students Able to Explain Genetic engineering
y Genetic engineering: y Direct manipulation on organism·s

genetic material in a way that does not occur under natural conditions
y Goals of genetic engineering: ‡ Improve understanding of how genes

work ‡ Advance biotechnology³the manipulation of organisms to create products or cure diseases

y Process of Genetic Engineering
Isolation of the Genes






What is cloning???
y Cloning

processes used to create copies of DNA fragments (molecular cloning), cells (cell cloning), or organisms.

y Three types of cloning: 1. Gene cloning a process where fragments of DNA are

transferred from one organism to another, usually carried on a DNA vector. Microbes like bacteria are convenient carriers and hosts for cloning DNA. 2. Reproductive cloning 3. Therapeutic cloning
y Basic of gene cloning technologies

recombinant DNA

Reproductive and theraputic Cloning

y Recombinant

DNA (rDNA) technologies is a genetic engineering method on biomolecular level.

y Recombinant DNA

artificial DNA that is created by combining two or more sequences on DNA that would normally occur together through the process of gene splicing.

y Recombinant DNA technologies use enzymes that cleave or copy

DNA in living cells. The purified enzymes can perform DNA manipulations in vitro in recombinant DNA experiments.

y General Steps in rDNA procedures:

The DNA of interest that is to be transferred (also called foreign DNA, insert DNA, cloned DNA, or transgene) is obtained by first extracting the DNA from the organism and then cutting out the specific DNA sequence using special enzymes. 2. The transgene is inserted into a special DNA molecule called a cloning vector and joined (by ligation) to produce a new recombinant DNA molecule (also called cloning vector-insert DNA construct, or simply DNA construct). 3. The DNA construct is transferred into, and maintained in, a host cell (bacterium) by the process of transformation. The vector replicates, producing identical copies (called clones) of the insert DNA.

The host cells that have incorporated the foreign DNA are identified and isolated from untransformed cells. 5. The cloned DNA can be manipulated such that the protein product it encodes can be expressed by the host cell.

y Vectors are entities for carrying the target DNA into a host y o o o

cell for multiplication or cloning. Cloning vectors: Plasmid vectors Viral vector Vectors for very large DNA fragments:
o Cosmids (hybrid of plasmid and bacteriophages) o Bacterial artificial chromosomes o Yeast artificial chromosomes o Shuttle vectors

a vector (usually a plasmid) constructed so that it can propagate in two different host species

Viral vectors (Phage)

Plasmid from E.coli

Cosmid vector

Stem cells
y Stem cells

special cells that have the ability to divide for an indefinite period and can give rise to a wide variety of specialized cell types.

y Types of stem cells based on its ability to differentiate: ‡ Totipotent stem cells differentiate to all kind of cells and able

to form new organism ‡ Pluripotent stem cells differentiate to derivates of three embryonal layer (ectoderm, mesoderm, endoderm) ‡ Multipotent stem cells differentiate to one specific organ or tissue ‡ Unipotent stem cells can·t differentiate to specific tissues, but have fast regeneration and proliferation abilities

y In human, there are two types of stem cells:

1. Embryonic stem cells 2. Adult stem cells

7. Students Able to Explain Ethical Aspects of Genetic Engineering
y Producing organisms with

y Fertile organism (ex:

desired phenotypes and genotypes y Supporting health and medical world (ex: producing insulin) y Reducing pollution y Producing efficient foods

watermelon) y May be used for negative purposes (ex: forming a battalion of army with cloned human) y Crisis of Identity y Against the religion

8. Students Able to Explain DNA Test
y What make we look different one another? y Junk DNA loci of STRs (Short Tandem Repeats) y STRs are sections of DNA arranged in back-to-back repetition (a y simple sequence is repeated several times in a row). y Example of STRs: y TCAT TCAT TCAT TCAT TCAT TCAT y The number of repeats found in pairs of STRs varies from one

person to another. STR DNA fingerprint of a person is completely different one another. The probability of anyone else having identical alleles with a person at each and every one of his or her loci is outrageously small.

General Process of DNA TEST
Collecting Biological Samples Extracting DNA Genetic Analysis

Collecting BIOlogical samples
y Common samples for DNA tests: ‡ Saliva ‡ Buccal swabs ‡ Semen ‡ Blood ‡ Hair

DNA Extractions
y General methods:

1. Break open the sample cells to free the DNA from the

nucleus (cell lysis). 2. Remove the proteins (which make up most of the biological sample) by digesting them with an enzyme. 3. Remove the DNA from the solution by adding alcohol.

y Using PCR (Polynucleotide Chain Reactions)

copies of

specific fragments of DNA molecules
y Why we need copies of DNA? ‡ Current technology used in DNA fingerprinting can·t detect

the DNA unless large amounts are present. ‡ Matches must be exact when it comes to DNA fingerprinting and forensic genetics. To avoid misidentifications, many STR loci from each sample must be examined.

PCR Methods

y Next step: y Read DNA fingerprint using electrophoresis

DNA molecules is negatively charged. So, electrophoresis separate the DNA fragments based on its lengths and sizes.

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