BIOTECHNOLOGY

APPLICATIONS IN HEALTH & HUMAN WELFARE

Dr. B. Siva Kumari Head.Dept of Botany. Andhra Loyola College

BIOTECHNOLOGY
The term coined by karl Erkey-1919
Combination of 2 words- biology+ technology The term is recent, but technology is prehistoric The technology is as old as human Our grand mother is a good technologist Technology boosted in 1970s after the discovery of DNA and GENE.

Definition
The technology involved in the utilization of living organisms in the production of useful products. It is a emerging branch in research It is used in the human welfare, health, industry, agriculture and protection of environment. It is also used in mining, fuel technology, food industry, pollution control.

Eukaryotic Chromosomes
Located in the nucleus Each chromosome consists of a single molecule of DNA and its associated proteins The DNA and protein complex found in eukaryotic chromosomes is called chromatin 1/3 DNA and 2/3 protein Complex interactions between proteins and nucleic acids in the chromosomes regulate gene and chromosomal function

Prokaryotic chromosomes
Circular double helix Complexed with protein in a structure termed the nucleoid Attached to plasma membrane

Karyotype
The representation of entire metaphase chromosomes in a cell, arranged in order of size and other characteristics

Ideogram
Diagramatic representation of a karyotype Individual chromsomes are recognized by -arm lengths p, short q, long -centromere position
metacentric sub-metacentric acrocentric telocentric

-staining (banding) patterns

From Miller & Therman (2001) Human Chromosomes, Springer

Facultative heterochromatin
All types of sequences C-banding negative Late replicating Condensed conformation Not transcribed Includes genes silenced in specific cell types and/or at specific times in development e.g. inactivated X chromosomes

Telomeres
End structures of linear chromosomes Serve to replicate chromosome ends Serve to stabilize chromosome ends (i.e. prevent nonhomologous end joining, NHEJ) G-rich tandem repeats - TTAGG, insects - TTAGGG, vertebrates - TTTAGGG, plants Length is under genetic and developmental control - e.g. 2-5 kb in Arabidopsis, 60-160 kb in Tobacco, 15 kb in humans Sequence and proteins conserved across taxa, mammals to plants

Centromeres
Primary constriction Kinetochore - spindle fiber attachment Region of sister chromatid cohesion Constitutive heterochromatin Repeat sequences - CENs - 5 to 170 bp e.g. human alphoid satellite repeat No universal centromere repeat, but the same repeat can be found in more than one centromere of a species or between species Centromere repeats can change rapidly in evolution via mutation, new elements, recruitment of other genomic repeats Specific associated proteins e.g. Centromere-specific histone HE (CenH3)

3. The form of eukaryotic chromosomes changes through the cell cycle:

a. In G1, each chromosome is a single structure b. In S, chromosomes duplicate into sister chromatids but remain joined at centromeres through G2 c. At M phase, sister chromatids separate into daughter chromosomes

4. In G1 eukaryotic chromosomes are linear dsDNA, and contain about twice as much protein as DNA by weight. The DNA-protein complex is called chromatin, and it is highly conserved in all eukaryotes.

Chromatin Structure
1. Both histones and non-histones are involved in physical structure of the chromosome. 2. Histones are abundant, small proteins with a net (+) charge. The five main types are H1, H2A, H2B, H3 and H4. By weight, chromosomes have equal amounts of DNA and histones. 3. Histones are highly conserved between species (H1 less than the others). 4. Histones organize DNA, condensing it and preparing it for further condensation by nonhistone proteins. This compaction is necessary to fit large amounts of DNA (2m/6.5ft in humans) into the nucleus of a cell. 5. Non-histone is a general name for other proteins associated with DNA. This is a big group, with some structural proteins, and some that bind only transiently. Non-histone proteins vary widely, even in different cells from the same organism. Most have a net (-) charge, and

6.Chromatin formation involves histones, and condenses the DNA so it will fit into the cell. Chromatin formation has two components: a. Two molecules each of histones H2A, H2B, H3 and H4 associate to form a nucleosome core, and DNA wraps around it 1 34 times for a 7-fold condensation factor. Nucleosome cores are about 11 nm in diameter

Fig. 2.24 Basic eukaryotic chromosome structure

b. H1 further condenses the DNA by connecting nucleosomes to create chromatin with a diameter of 30nm, for an additional 6 fold condensation. The solenoid model proposes that the nucleosomes form a spiral with 6 nucleosomes per turn (Figures 2.24 and 2.25).

Fig. 2.25 the 30-nm chromatin fiber

Peter J. Russell, iGenetics: Copyright Pearson Education, Inc., publishing as Benjamin Cummings.

Centromeric and Telomeric DNA

1. Centromeres and telomeres are eukaryotic chromosomal regions with special functions. 2. Centromeres are the site of the kinetochore, where spindle fibers attach during mitosis and meiosis. They are required for accurate segregation of chromatids. 3. Yeast (Saccharomyces cerevisiae) centromeres are well-studied. Called CEN regions, their sequence and organization are similar, but not identical, between the chromosomes. Other eukaryotes have different centromere sequences, so while function is conserved, it is not due to a single type of DNA sequence. (Fig.

The genomic code for nucleosome positioning

DNA double helix Nucleosomes

Chromosome
Felsenfeld & Groudine, Nature (2003)

Isolation of natural nucleosome DNAs

Digest unwrapped DNA

Extract protected DNA Clone, sequence, analyze individuals