B.Tech Project (2012-2013)

Mr. Rex Arunraj, Genetic Engineering Lab, Dept of Genetic Engineering
DNA Barcoding of Plant Powder
Number of students: 2

DNA barcoding is identifying an species or an adulterant using DNA barcodes. The project involves DNA isolation from plant powders and PCR amplification of the same. The amplified product will be investigated for the species or authenticity.

Dr. Devi.A, Stemcell Biology Lab, Dept of Genetic Engineering
Study on the expression of Notch receptors in human gastric cancer
Number of Students:2
Gastric or Stomach cancer is the second leading cause of cancer death globally with more than 700,000 deaths each year, and is particularly common in East Asia. Treatment of this deadly disease is often difficult and unsuccessful because of the asymptomatic nature of the disease and poor understanding of the cause. Until now, the molecular mechanisms that cause stomach cancer remains a mystery. In search of better therapeutic options for stomach cancer, Notch signaling has emerged as a potential target. Notch signaling is an important molecular pathway involved in the determination of cell fate. In recent years, this signaling has been frequently reported to play a critical role in maintaining progenitor/stem cell populations as well as a balance between cell proliferation, differentiation and apoptosis. Thus, Notch signaling may be mechanistically involved in carcinogenesis. Of the various pathways in gastric cancer, it is known that deregulation of Notch pathway plays a vital role in tumorogenesis and therefore a better understanding of the target genes involved is crucial for the development of new therapeutic drugs.
Recent studies on the several gastric cancer cell lines predicted that the NOTCH1, NOTCH2 and NOTCH3 were expressed in all the gastric cancer cell lines. Still, the gene expressions on the gastric cancer tissue samples remain questionable and this work aims to analyze the expression of Notch receptors that are predominantly expressed in different stages of gastric cancer by Immunohistochemistry, western blotting and RT-PCR. 

Dr. Usha, Genetic Engineering Lab, Dept of Genetic Engineering
Role of rice sucrose transporter genes in root exudation
Number of Students: 4

Root exudates are the chemicals secreted into the soil by roots which regulate the soil microbial community in their immediate vicinity. Root exudates give resistance to pathogen, combat pathogenic micro-organisms, and encourage beneficial symbioses by attracting beneficial micro-organisms. The SUT genes if highly expressed may affect the sucrose exudation level in the roots. Comparison of the differences between the sucrose exudation profile of the wild type tobacco plant and that of overexpressed transgenic tobacco plants can be done to check this hypothesis. Investigation of the sucrose exudation profile in relation to SUT genes of rice can be done by over-expressing these genes in the tobacco plants. Further investigations can be done by gene knockout of tobacco plants’ SUT genes highly homologous to rice sucrose transporter genes and then extrapolated to rice sucrose transporter genes to elucidate the key functions of rice SUT genes. And finally the reverse transcription of all of the rice sucrose transporter genes and then check the sucrose exudation level via roots with the help of HPLC method. The use of fertilizers has resulted in loss of plants’ ability to acquire nutrients. SUT genes analyses/overexpression can help increase plant performance, resistance and nutrient activity. This can further help in elucidation of increased plant performance, altering role of root exudates, by using gene responsible for particular exudates in economically important crops.

Mr. S. Iyappan, Molecular Genetics Lab, Dept of Genetic Engineering
Mutation analysis of Morquio syndrome
Number of students: 2 

Morquio syndrome is an autosomal recessive trait. That means both your parents must pass you the defective gene in order for you to get this disease.  There are two forms of Morquio syndrome: Type A and Type B.
  • Persons with Type A do not have a substance (enzyme) called galactosamine-6-sulfatase.
  • Persons with Type B do not produce enough of an enzyme called beta-galactosidase.
The body needs these enzymes to break down a long strand of sugar molecules called the keratan sulfate sugar chain. In both types, abnormally large amounts of glycosaminoglycans build up in the body and brain, which can damage organs.  The syndrome is estimated to occur in 1 of every 200,000 births. Symptoms usually start between ages 1 and 3. A family history of the syndrome raises one's risk for the condition.
This study will be conducted to identify mutation that could be used to molecular diagnosis of the disease and genetic counseling for the family cases with accumulation mucopolysaccharidosis (MPS) (Morquio syndrome IVA).  Collection of blood sample form the patient with morquio syndrome IVA, isolation of DNA, PCR amplification of the galactoseaminesulphate sulphatase gene exons, preliminary analysis with SSCP, sequencing of the mutated exons. The samples will be collected from the suspected carrier in the family members based on pedigree analysis.

Mr. Rathinasabapathi, Genetic Engineering Lab, Dept of Genetic Engineering
Cloning of T3SS effectors from Pseudomonas syringae
Number of students: 3

 Plant associated organism’s secrete protein to modulate the plant defense circuitry. These proteins are known as effectors and these effectors help the pathogen for its infection by suppressing the plant basal defense mechanism. Pseudomonas syringae is a plant pathogen which secretes different kinds of effector proteins to enhance its virulence. Since these effectors suppress the plant immunity, it may be useful to enhance the expression of heterologous genes in transient expression system. In this study, the genes coding for T3SS effectors would be amplified from the P.syringae and cloned into suitable vectors. 

Mr. N.Manojkumar, Bioseparation Lab, Dept of Genetic Engineering
Cloning of a Fibrinolytic Enzyme Gene from Bacillus Species in Escherichia coli.
Number of Students: 2
Several investigations are being pursued to enhance the efficacy and specificity of fibrinolytic therapy. In this regard, microbial fibrinolytic enzymes attracted much more medical interests during these decades. Fibrinolytic enzyme is quite common in Gram-positive bacteria, and Bacillus species stand out in particular, as many extracellular and even intracellular variants have been identified.  The present work, the fibrinolytic gene from Bacillus species will be cloned into the pET vector and expressed in Escherichia coli strain. Total genomic DNA will be isolated and by using specific primers PCR amplification of the fibrinolytic gene will be done. SDS-PAGE and enzyme assay will be done for characterizing the expressed protein. The DNA and amino acid sequence alignments will be done using BLAST search. This study may provide evidence that fibrinolytic protein can be actively expressed in E. coli. The commercial availability of fibrinolytic enzyme is of great importance for industrial applications and also pharmaceutical purposes as thrombolytic agent. Thus, the characterization of new recombinant fibrinolytic enzyme and the development of rapid, simple, and effective production methods are not only of academic interest, but also of practical importance.

Mr. N.Aruljothi, Bioseparation Lab, Dept of Genetic Engineering
Human LDLR gene mutation analysis in Myocardial Infarction patients in South India
Number of Students: 2
Accumulation of low density lipid in the blood vessels is one of the causes for Myocardial Infarction. This condition may occur due of mutations in LDL receptor gene, which leads to failure in uptake of LDL by hepatic cells and leads to increased levels of LDL in blood. There are more than 1000 mutations known to occur in LDLR gene. The study involves screening of LDLR gene for mutations in Young Myocardial Infarction patients all over South India. 

Dr.K.N.Rajnish, Genetic Engineering Lab, Dept of Genetic Engineering
Cloning and expression and of a putative genes from Zymomonas mobilis ZM4
Number of students: 3
Zymomonas mobilis is an ethanologenic bacterium that ferments simple sugars to produce ethanol. This bacterium is well suited for industrial applications due to advantages such as high rate of sugar uptake and ethanol tolerance. The genome sequencing project of three species of Zymomonas has indicated the presence of genes with putative function. In this context, three putative genes coding for an aminopeptidase (pepN), beta-galactosidase (bga) and metallo-betalactamase (bla) were selected for detailed analysis. Sequence and homology based analysis would be carried out to detect the presence of homologues and conserved domains. The ORFs will be PCR amplified and cloned into expression vectors. The recombinant protein would be purified and would be assayed for the predicted function.

Dr.R.Srividhya, Genetic Engineering Lab, Dept of Genetic Engineering
Isolation and purification of the enzyme acetylcholine esterase and studies on in silico interactions with neuroprotective agents
Number of students: 3
Acetylcholine esterase is an enzyme that is present in the neuromuscular junctions useful for the recycling of choline to the presynaptic terminals. Studies on the enzymes involved in neurotransmission at the synaptic junctions will give us an idea about the cognitive status of the animal/subject under study. Isolation of the enzyme from rat brain tissue will be carried out using salting out (ammonium sulphate saturation), ion exchange chromatography and electrophoresis. Enzyme kinetic studies will be carried out. Interaction of the enzyme with several neuroprotective agents could be compared with that of the enzyme’s own inhibitor and substrate. Molecular docking studies will also be carried out using bioinformatic tools like AutoDock 4.0 to cross-check the in-silico interactions.Similar studies can be carried out for other enzymes like Choline acetyltransferase involved in the cholinergic system or monoamine oxidase involved in dopaminergic system.

Dr.N.S.Raja, Genetic Engineering Lab, Dept of Genetic Engineering
Study of changes in the metallobiomics and gene expression patterns of rice and tobacco roots in response to Cr(III) and saline stress
Number of students: 3
Transcriptional regulation, also known as transcriptome reprogramming, is an important way by which plants adapt to biotic and abiotic stress. Study of physiological and molecular effects of environmental stress will lead to the identification of genes involved and their expression patterns during the course of stress perception and response by the plants. The molecular mechanisms that are involved in different abiotic stress have been revealed independently in few cases. Understanding of convergence points between different stress signalling pathways remain rudimentary. Therefore, it is necessary to explore the cross talk between the different stress responses. High-throughput genomic technologies have made it possible to analyze expression of thousands of genes at a time. Cr(III) and NaCl are the two major pollutants of tannery Profiling of different genes expressed under Cr(III) and saline stress individually will identify the genes responsible for tolerance, accumulation and defense response in plants with respect to Cr(III) and saline stress. Cross talk between two different abiotic stress will be analysed by subjecting the plants to combined Cr(III) and saline stress. This system will mimic the tannery effluent polluted environment where plants are exposed to saline and chromium sterss. This study will help to develop transgenic plant capable of surviving in tann.

Mrs. S. Rekha, Genomics Lab, Dept of Genetic Engineering
Case - Control study of Abra1 gene in South Indian Population
Number of Students: 2

Breast and ovarian cancers are the most common type of cancers affecting women worldwide. According to AIIMS, Delhi, the rate of breast and ovarian cancers in Indian metro cities is likely to increase from 21.7 to 28.7 percent by 2015. A recent study on Finland population showed mutations in Abra1 gene, coding for a protein Abraxas involved in BRCA complex for DNA repair mechanism. We are screening for common variants in Abraxas protein in clinically proven breast and ovarian cancer patients among south Indian population. To study the occurrence of the identified variants in population, a case control study has to be carried out for the variants. The exons of control (Negative breast cancer) will be amplified and analysed for identified variants by SSCP and confirmed by sequencing.