Biotech : Mol-Sel Laboratory [PDF]

Animal-Cloning). Technical Aspect: Part 2. GC. Class Discussion. 6. Software analysis of Chromosome. Cells. GC+ Lab. Tec

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Idea Transcript


No

Subject : Cellular

Lecturer

Note

1.

Introduction-Recent dev of . Anim Biotech:Mol-Cell

GC

Class Discussion

2

Basic Mol-Cell Laboratory Practice, In vitro.

GC

Class Discussion

3

Chromosome Bio-Cell Analysis

3.

In vitro- Cell culture (IVM, IVF, IVG)

GC

Class Discussion

4.

Nuclear Cell – Gene (Transgenic Animal-Cloning: Cell communication and Cell Reprogramming) Basic Theory : Part 1

GC

Class Discussion

5.

Nuclear Cell – Gene (Transgenic Animal-Cloning) Technical Aspect: Part 2.

GC

Class Discussion

6.

Software analysis of Chromosome Cells

GC+ Lab. Tech.

Lab works/Visit/LSIH

7.

Nuclear Cells Transfer /Gen Transfer

GC + Lab. Tech

Lab work/Visit/LSIH (Jika Lab-tidak sedang

Biotech : Basic Mol-Sel Laboratory- in Vitro •Most widely used biotech products are recombinant proteins (produced by gene cloning in cell culture) •Cell culture refers to the technique of growing cells in a lab under controlled conditions; similar to in vitro •“in vitro” refers to working in a controlled environment outside of a living organism •Bioreactors (large culturing “containers” where DNA of interest can be mass produced) are also used

Animal Cell Biotechnology: Desinfection step

Tissue isolation

Incubation&growth

Primary cells

Animal Cell Culture: Biotechnology

Donor cell

Remove nucleus Add somatic cell from adult donor from egg cell

Nucleus from donor cell

Implant blastocyst in surrogate mother

Clone of donor is born (reproductive cloning)

Remove embryonic stem cells from blastocyst and grow in culture

Induce stem cells to form specialized cells (therapeutic cloning)

Grow in culture to produce an early embryo (blastocyst)

In vItro Technology: pH, temperatur, C02/O2, etc.

Kultur Sel : 1. In Vitro Maturation (IVM) of Oocyte 2. In Vitro Fertilization and Development 2. In Vitro Growth (IVG) : Kultur Folikel Memahami : Cell development (folikulogenesis)

Material : Oocyte.-Cumulus-Granulosa Cell Complexes from large antral follicles (4 – 6 mm) Medium : TCM199, (10 % FCS, 0.1 ng/ml Na-

pyruvate, Antibiotics. Hormon : FSH and LH Temperatur

: 38.5 – 39 0C

Gas %

: 5 % CO2 in air

Dish

: Falcon 35 mm

Drop

: 100 ul ( 10 ul/oocyte), covered with mineral oil/ parafin oil

Cellular Analysis : Karyotyping

Meiosis KM

6

Identifikasi Gen Pada posisi Kromosom Terapi Gen

Biotech Treatments •In the near future, it may be commonplace for treatments to include the use of gene therapy (attempt to replace “defective” gene with “normal” gene) and tissue engineering (designing & growing tissues for use in regenerative medicines).

•1st Genetically Modified Organism (GMO) to produce human protein was E. coli (pictured right) that was given DNA to produce somatostatin (hGH -human growth hormone 1977)

Molecular Pharming • Molecular pharming is the use of genetically modified plants (or animals) as a source of pharmaceutical products.

• These are usually recombinant proteins with a therapeutic value.

• This is an emerging but very challenging field that requires: •manipulation (at the genetic engineering level) of protein glycosylation (addition of polysaccharide chain) •subcellular protein targeting in plant cells

Animal Applications •Animals can be used as bioreactors! •Many human therapeutic proteins are needed in massive quantities (>100s of kgs), so scientists create female transgenic animals to express therapeutic proteins in milk.

• Goats, cattle, sheep, & chickens are sources of antibodies (protective proteins that recognize & destroy foreign material)

•Transgenic refers to containing genes from another source

Knock Outs • Basic research in biotech uses knock-out experiments, which are very helpful for learning about the function of a gene. • A knock-out is created when an active gene is replaced with DNA that has no functional information. • Without the gene present, it may be possible to determine how the gene affects the organism (its function)

Dolly In 1996, Dolly the sheep became the first cloned animal created by the somatic cell nuclear transfer process.

• Born: July 5, 1996 • Announced: February 22, 1997 • Died: February 14, 2003 • Dolly was cloned from a cell taken from a six-year-old ewe • She became the center of much controversy that still exists today

Forensic Applications •DNA fingerprinting is the classic example of a forensic application. It is used most commonly for law enforcement and crime scene investigation (CSI). •It was first used in 1987 to convict a rapist in England.

•Other applications of DNA fingerprinting include • identifying human remains • paternity tests • endangered species (reduces poaching)

• epidemiology (spread of disease )

Implementasi: Paternity Test (Human) • •

– DNA 

+

Mom

F1

Who’s the father? Ternak: Similaritas DNA

F2

child

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