ANNUAL REPORT - Department of Biotechnology

ATPase mRNA binding protein, cathepsin L and
trefoil factor 3 that play critical roles in cellular
processes were found to be significantly altered
during the process of aging. Promoters of iPPase, 3'
non-translated β-F1 ATPase mRNA binding protein
and cathepsin L gene were cloned and the location of
transcription start site of iPPase gene identified by
primer extension analysis. Sequencing of
senescence marker protein (SMP30) promoter up to
-3 kb and DNA-protein analysis have identified a total
of 30 nuclear factor binding sites within -2.5 kb to -
800 bp of transcription start site of which 10 sites
(Oct-1, GATA-1, GATA-2, C/EBP, CdxA, AP-1, Nkx-
2.5, SRY, c-Ets and Ik-2) have been confirmed by
EMSA. Age-dependent alterations in the activity of
transcription factors and androgen receptor
indicated that the transcription factors, e.g. SRY,
HSF2 and GATA have an important role to play in
aging. The other transcription factors such as HSF2,
Ikaros, GATA, Pbx, SRY were also found to be
involved in the differentiation and development of
organisms. The binding of these transcription factors
to the AR promoter which has age-dependent
expression strongly indicated a regulatory role
during senescence. C/EBP, CREB, AP-1, Oct-1, Sp1
and IRF also have been reported to play a regulatory
role in hepatic genes and may play a significant role
in age-dependent down regulation of rat androgen
receptor gene. The varying level of transcription
factors and their expression during aging indicate a
complex interplay of transcription factors regulating
the decline in AR expression during aging.
Molecular biology of Cancer
Studies on proto-oncogene EVI1 in a subgroup of
chronic myeloid leukemia patients progressing to
blast crisis were undertaken. Several deletion
mutants of EVI1 were constructed and P/CAF shown
to acetylate EVI1 at the proximal part of the protein.
Site directed mutagenesis revealed acetylation of
EVI1 at lysine 367 residues.
Infectious Diseases
A septaplex PCR assay was developed for rapid
identification of species-specific virulent and sxtpositive
strains of V. cholerae and one hundred
strains of V. cholerae O1 were tested to document
the validity of assay. PCR testing of Vibrio cholerae
DBT Annual Report 2006-07
192
O1 biotype El Tor serotype Inaba associated with an
outbreak of cholera revealed that all of the five
isolates examined carried the TCP pathogenicity
island, the CTX genetic element, the RTX toxin, and
produced cholera toxin (CT). Restriction fragment
length polymorphism (RFLP) analysis revealed that
these Inaba isolates possess a single copy of the
CTX element flanked by two tandemly arranged
copies of the RS element upstream of the core
region. The isolates were resistant to ampicillin,
nalidixic acid, trimethoprim, sulfamethoxazole,
streptomycin, and to the vibriostatic agent.
Ribotyping of these Inaba isolates revealed a
hybridization profile similar to a strain of serotype
Ogawa prevalent in Southern India.
Molecular ecology of malaria vectors were analyzed
by comparing sequences of rDNA of various
Anopheles species. Four novel sequences of ITS2
region of rDNA of Anopheles fluviatilis were
identified. A multiplex PCR assay to detect fluviatilis
sibling species developed during the course of the
year will be used to understand feeding habits
(Anthropophilic index) and sporozoite carrying
capacity of these vectors.
Bioprospecting of Microbes
Studies on bio-prospecting were continued with a
view to tap the vast potential of thermopiles. A diverse
group of bacteria belonging to the genera
Thiomonas, Comamonas and Chromobacterium
were isolated from previously unexplored hot
springs. A chemolithoheterotrophic, thiosulfate
oxidizing, gram negative bacterium (designated
strain S10) was isolated and identified. 16S DNA
sequence data and the total fatty acid analysis
suggested it to be a new species of genus
Thiomonas for which the name Thiomonas
bhubaneswarensis has been proposed. A mutant of
Mesorhizobium ciceri unable to grow on C4- compounds (succinate, malate and fumarate)
forming symbiotically defective nodule on the roots of
chickpea (Cicer arietinum L) has been identified.
Characterization of an rpoN deficient mutant
provided evidence that the rpoN encoded alternative
sigma factor is required for symbiotic function in M.
ciceri, suggesting that like Rhizobium spp. strains
NGR234 and Rhizobium etli, the rpoN encoded
alternative sigma factor of Mesorhizobium ciceri also
played a role in symbiotic nitrogen fixation.