5. Book 2:
Application of Biotechnology in Sericulture
Editor: R. Venkatesh Kumar, V. Shyam Kumar, K. Sashindran Nair, and C.K. Kamble
Publisher: Studium Press (India) Pvt Ltd
Available in Amazone !
7. Modern Biotech
• basic laws of heredity: mid 1800’s
• Findings were published in 1866 (ignored for 34
years)
8. Biotechnology
• "Biotechnology“: coined in 1919
by Karl Ereky, an Hungarian
engineer
• ‘the use of living organisms or
their products to modify human
health and the human
environment.’
Karl Ereky
9. • "Biotechnology“: coined in 1919 by Karoly (Karl)
Ereky, Hungary (‘the use of living organisms or
their products to modify human health and the
human environment.’)
• Frederick Griffith’s Mouse Experiment: 1928, UK
Modern Biotechnology
10. Oswald T. Avery’s Transformation
Experiment - 1944
Determined that “IIIS” DNA was the genetic material
responsible for Griffith’s results (not RNA).
11. Hershey-Chase Bacteriophage
Experiment – 1952-53
1. T2 bacteriophage is composed of DNA and proteins:
2. Set-up two replicates:
• Label DNA with 32P
• Label Protein with 35S
3. Infected E. coli bacteria with two types of labeled T2
4. 32P is discovered within the bacteria and progeny phages, whereas
35S is not found within the bacteria
Alfred Hershey
Martha Chase
12. DNA Replication
• Nucleotides always added to the growing strand at the 3‘
–OH
• The OH-group reacts with the phosphate group on the 5'
C of the deoxyribose so the chain grows
• Energy is released when the bond linking 2 of the 3
phosphate groups to the deoxyribonucleoside
triphosphate breaks
• Remaining phosphate group becomes part of the sugar-
phosphate backbone
13. DNA Replication: features
• Semi-conservative
• Semi-discontinuous
• DNA-bases come with their own energy source
• Starts at ‘Ori’
• Bi or Uni-directional
• 5’ to 3’ direction
• Need RNA
14. an experiment to prove that DNA replication was semiconservative.
Semiconservative replication means that when the double stranded DNA helix
was replicated, each of the two double stranded DNA helices consisted of one
strand
Nitrogen is a major constituent of DNA, the genetic material of all cells.
It is commonly found in the 14N isotope, but it can also be found in the heavier
15N isotope.
Meselson-Stahl
Experiment,
1958
16. Rosalind Elsie Franklin
• mid 1900’s
• Led to discovery of
structure of DNA – the
double helix
Physical basis of heredity : the DNA
17. • James Watson
• Francis Crick
• Collaborated to produce the first
model of DNA structure in 1953
James Watson
Francis Crick
Physical basis of heredity : the DNA
18. •Sir Louis Pasteur in 1870 discovered that pebrine
disease goes from mother to offspring.
Most seri-biotechnology progress
made in Japan.
•Breeding of silkworm was highly frequent in Japan in early 20th
century.
•Hybrid vigour was shown in 1906 by Toyama.
•The W-chromosome was shown to be involved in sex
determination by Hasimoto (1933).
•Virus resistance against NPV, CPV and IFV have been reported;
they are polygenic characters (Aratake, 1973).
•The fibroin heavy chain (Fibroin H) gene from B. mori was
isolated by Goldsmith, 1984.
History of seri-biotechnology
19. (1) A large number of expressed sequence tag (EST) sequences for
silkworm multiple tissues have been sequenced and released in
2002.
(2) The draft genome sequences were independently generated by
Chinese and Japanese groups in 2004 (Xia et al. and Mita et al.).
(3) Finally, Chinese and Japanese groups worked jointly and assembled a
high-quality genome sequence of the silkworm in 2007.
The silkworm genome project
20. Silkworm is the first lepidopteron to have its genome
mapped.
• Genome: ~ 530 MB
• They identified five key genes relating to gender control,
growth and immunity.
• They also found 1874 genes associated with the silk
gland. Silkworm has an estimated 18,510 genes.
21. • Production of biomolecules in silkworm: expression of alpha-
interferon and interleukin-3in silkworm was shown in 1985
(Maeda).
• Transposon like elements discovered in silkworm DNA in 1990
(Michaille).
• Transcriptional gene regulation was shown in 1990s.
• Transgenic silkworm generated experimentally (Tamura, 2000).
• Tissue culture of silkworm brain done in RRL, Jorhat (Das, 2004).
• PGPR applied for silkworm host improvement (Unni, 2008; RRL,
Jorhat). They also tried biocontrol of silkworm diseases. They
have also tried indegenous medicinal plants to control silkworm
diseases.
Few recent achievements in seri-
biotechnology
22. Morus notabilis genome database, 2013
(by Wang, BGI; Ji, Sericultural Research Institute and Xiang, Southwest
University)
23. •Seri Biotech Research Laboratory, established by Central silk board (CSB, 1948)
at Bangalore in 1993.
•Identification of DNA marker for NPV resistance in silkworm (CDFD, Hyderabad
and CSB).
•Genetic linkage map for the silkworm established employing 518 simple
sequence repeat or microsatellite markers (Nagaraju, with China, Japan
and USA; 2005).
•High yielding productive silkworm hybrids (viz., Swarnandhra, Hemavathy and
Kalpavatharavu) developed using DNA marker technology and released
to farmers of Andhra Pradesh, Karnataka and Tamil Nadu (2002).
•Luciferase and green fluorescent proteins were successfully expressed utilizing
silkworm as a bioreactor (Indian Institute of Science, Bangalore; 1998).
•Process for degumming of silk with fungal protease enzyme has been
standardized and found viable (IIT-Delhi, 2000).
•Mapping populations of mulberry for water use efficiency and genotyping of
trait-specific mapping population of mulberry (2013-2014).
•Transgenic mulberry lines with HVA-1 gene for abiotic stress tolerance have been
developed (University of Delhi South Campus, 2012).
History of seribiotechnology in
India
24. Central Sericultural Research and Training Institute, Mysore
(Estd. 1961)
Silk Protein Database
Silk transcription factor Database
Silk protein-protein database
25. •Baculovirus-resistant silkworm generated through RNAi technology
- 2013.
• Potential immune-related genes discovered -2006.
• Eleven potential silkworm (Bombyx mori) lines tolerant to BmNPV
developed 2015-16.
• Two genes, possibly involved in the silkworm sex determination,
were identified.
• Consortium Project on Mulberry Genomics (CPMG) has been
developed in Morus alba.
• Transcriptome analysis has been carried out in Mulberry.
Silkworm biotechnology - present research in India
26. Silkworm biotechnology – commercial approaches
EntoGenetics (USA): transgenic silkworm to produce spider silk (it is
stronger).
Kraig Biocraft Laboratories (USA): transgenic silkworm to produce
composite silk (silkworm silk and spider silk). Founded in 2006.
AMSilk (Germany) & Korea Advanced Institute of Science & Technology
(KAIST) (South Korea): spider silk through a genetically engineered E.
Coli bacteria. AMSilk is using spider silk in repair and for drug
delivery.
Araknitek(USA), Entogenetics (USA): the most innovative approach.
Started by Nexia Biotechnologies, to breed “spidergoats”—goats
that could produce the silk proteins in the animals’ milk (called
Biosteel).
Araknitek, is also pursuing other methods to boost production of
the spider proteins, including transgenic alfalfa, bacteria, and
silkworms.
