Production of tENDO1 in stably transformed tobacco cell cultures for mismatch detection.

Scanning DNA sequences for polymorphisms and mutations often involve the mismatch specific cleavage by endonucleases at the mismatch sites and subsequent analysis of the digested product for mutation discovery. One of the limitations of using enzymatic mutation detection methods are the cost and availability of a mismatch specific endonuclease. We report the establishment of Nicotiana tabacum L. Cv. Bright Yellow 2 cells stably expressing the truncated ENDO1 (tENDO1) mismatch specific endonuclease. The 5′-Untranslated region of N. tabacum alcohol dehydrogenase gene (NtADH 5′-UTR) under the control of cauliflower mosaic virus (CaMV 35S) promoter was employed to improve the tENDO1 protein yield. To ease the purification process, tENDO1 was secreted into the culture medium and isolated using nickel affinity chromatography. The tENDO1 was estimated to be stably produced in an average of 0.7–0.9 % total soluble protein. Functional test on tENDO1 for mismatch detection demonstrated that tENDO1 retained mismatch specific endonuclease activity resembles its native protein. Further biochemical analysis showed that tENDO1 exhibited mismatch detection specificity and efficiency comparable to other commonly used endonucleases

Newcastle disease virus: macromolecules and opportunities

Over the past two decades, enormous advances have occurred in the structural and biological characterization of Newcastle disease virus (NDV). As a result, not only the complete sequence of the viral genome has been fully determined, but also a clearer understanding of the viral proteins and their respective roles in the life cycle has been achieved. This article reviews the progress in the molecular biology of NDV with emphasis on the new technologies. It also identifies the fundamental problems that need to be addressed and attempts to predict some research opportunities in NDV that can be realized in the near future for the diagnosis, prevention and treatment of disease(s)

Newcastle disease virus: a journey from poultry to cancer

Infectious diseases have had a great impact on human lives and civilizations since time immemorial. Success in controlling and eradicating these diseases, on the other hand, has not only overcome human misery but expedited human progress, both individually and collectively. The eradication of smallpox and the successful control of poliomyelitis and hepatitis B can even be measured in terms of the improvement in the world’s GDP. However, new and re-emerging infections, including those connected to livestock such as the Nipah virus, SARS and avian influenza, remind us to be ever vigilant and responsive to new threats. The Newcastle disease virus (NDV) causes a highly contagious and fatal respiratory disease in chickens and other types of birds and affects the poultry industry worldwide. Although this virus is effectively controlled by vaccination, it is still endemic in Malaysia and outbreaks of the disease have contributed to major losses for the poultry industry in the country in terms of mortality and loss in egg and meat production. By delineating the genome of this enveloped negative single-stranded RNA virus, various genes have been sequenced, cloned and expressed in plasmid vectors in various organisms. Knowledge of the structural and biological characterization of NDV has provided insights into various applications of this virus in the prevention and treatment of NDV and other diseases as well. Although the virus replicates poorly in normal human cells, causing mild respiratory disease and conjunctivitis, it replicates very well in human cancer cells. This provides a potential for its use in oncology. Cancer is an omnipresent threat to human lives, especially with, but not limited to, ageing, a worldwide modern phenomenon. Its control and eradication remain largely elusive, requiring considerable concerted efforts from everyone. The oncolytic activities of the virus against human malignancies lead to tumor apoptosis and cell death. In this respect, work is in progress to leverage NDV as a viral vector to express and deliver foreign genes for vaccination and gene therapy purposes for specific malignancies in humans. This of course opens up a whole new vista in research and industry, stretching imaginations and human potentials farther, with the cherished hope of improving the lives of human beings further

Peptide inhibitors against influenza virus

Influenza A virus is a particularly problematic virus because of its ability to cause high levels of morbidity on a global scale within a remarkably short period of time. It also has the potential to kill very large numbers of people as occurred in the Spanish influenza pandemic in 1918. Options for antiviral therapy are limited because of the paucity of available drugs and the rapid mutation rate of the virus leading to the emergence of drug-resistant strains. The current H1N1 pandemic and potential threats posed by other strains highlight the need to develop novel therapeutic and prophylactic strategies. Here, we summarize the current state and recent developments of peptide-based inhibitors of influenza A virus

Identification of Phosphoprotien : Phosphoprotien and Phosphoprotien : Nucleocapsid Protein Interaction Domains of New Castle Disease Virus

Summary : The yeast two hybrid system has been used to identify domains of the Newcastle disease virus ( NDV ) phosphoprotien (P) involved in self –association and interaction with the nucleocapsid protein (NP). Deletion analysis was used to map the domain (s) of the P protein involved in P:P and P:NP interactions. The C- terminal 45 amino acids (residues 247-291) were shown to play a major role in both of interaction. Comparison of these finding with other report suggests that paramyxoviruses are different with respect to interaction domain(s) between these two essential viral proteins involved in genome replication

First report of tomato mosaic tobamovirus from Malaysia

A mosaic disease of tomato was observed in Cameron Highlands, Malaysia and a tobamovirus was implicated as the cause based on virus particle morphology and reproduction of symptoms in Lycopersicon esculentum Mill. The virus was identified as tomato mosaic tobamovirus (ToMV) based on host range and serological properties

A General Survey of Nitrate-Nitrogen Levels In Well-water under Different Landuses

A general survey on N03 - - N levels in well-water under different landuses was conducted. Four areas were selected to represent different landuses namely mixedfarming (crop and animal production), forest, urban, and crop production only. Water samples from a total of 22 wells in the selected areas were collected and analysedfor N03- - N. The field and statistical data showed that the differences in N03 - - N levels in well- waterfrom different landuses were significant except for agriculture + animal farming. The highest N03 - - N level was observed in agricultural areas followed by urban and forested areas

Detection and precipitation of hepatitis B core antigen using a fusion bacteriophage

The nucleocapsids of hepatitis B virus (HBV) are made of 180 or 240 subunits of core proteins or known as core antigens (HBcAg). A fusion bacteriophage bearing the WSFFSNI sequence that interacts tightly to HBcAg was employed as a diagnostic reagent for the detection of the antigen using the phage-enzyme-linked immunosorbent (phage-ELISA), dot blot and immunoprecipitation assays. The results from phage-ELISA and dot blot assay showed that as low as 10 ng of HBcAg can be detected optimally by 1.0×1012 pfu/ml fusion M13 bacteriophage. The sensitivity of the dot blot assay corresponds with that of the phage-ELISA. HBcAg in HBV positive serum samples can also be detected using the fusion phage via the phage-ELISA and phage-dot blot assay. The phage cross-linked to cyanogen bromide (CNBr) activated agarose can also be used to precipitate HBcAg in bacterial lysate. The optimum amount of phage needed for cross-linking to 1 g of agarose is about 7.0×106 pfu/ml which could also precipitate purified and unpurified HBcAg in bacterial lysate. This study demonstrates the potential of fusion bacteriophage bearing the sequence WSFFSNI as a diagnostic reagent and a ligand for the detection and purification of HBcAg respectively