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Baculovirus is a host specific insect pathogen, which is characteristically identified by arrested host cell cycle on infection. This is a study experiment which investigates how the virus achieves its effects of cell pathology (CPE) [url=http://www.maximoupgrade.com/hot.php]hollister[/url] on the host insect cells such as vacuolation, polyhedron, cell enlargement and granulation on its transfection and infection. An effective transfection procedure for host SF9 (Spodoptera Frugiperda) cells is analyzed on gel electrophoresis technique to observe effects on [url=http://www.gotprintsigns.com/monclerpascher/]moncler[/url] SF9 cells (Dwight, 2010). This process use a gel agarose to separate DNA fragments on the basis of their molecular sizes and visualize them through a U.V. (Ultra-Violet) light trans-illuminator after a fluorescent dye staining (such as ethidium bromide).
The study experiment demonstrates that there is an indication of cell pathology (CPE) in the positive control and transfected cells unlike the normal cell structure in the negative control where there was no introduction of the baculovirus genome AcNPV (Autographa californicanuclear polyhedrosis virus). The conclusion obtained is that a transfection of a baculovirus DNA to host insect cells leads to infectious baculovirus progeny in SF9 cells.
Baculovirus is a viral pathogen which enters the host system of certain insects, leading to an ultimate cause an illness through generation of cell pathology. The virus infects specific populations of insects and this has led to their extensive use as selective biocide on target pesticides over a long period of time (Mohan and Gobinathan, 1992).
The specificity of these viral infection to insects has encouraged research studies to facilitate genetic modification of the viral genome so as to obtain virulence for certain targeted host. Since frequent use of chemical pesticides in our modern farming has been attributed as an important danger exposure to human and animal health, development of biological pesticides is highly advised. This is a significant factor for research studies formulated to analyze infection trends of biological pathogens such as baculoviruses on host insects to be developed.
These obligate infectious viruses are genetically coded by rod shaped neucleocapsids, which exists in the form of a double stranded, circular, super coiled gene, surrounded by a polyhedron matrix. Baculovirus infection in insects leads to an induction of membrane bound structures within the host cell nucleoplasms thus subsequently interruption of normal cell physiology. The biochemical explanation of how these membrane structures are formed and regulated is insufficient; however, cell cycle is suggested to have an influential role in the regulation of these intranuclear membranes on the basis of the normal dynamics of cellular membranes and the nuclear envelope model.
Surrounding the embedded virions this polyhedron matrix helps in protection of genetic material (viral DNA) until they are introduced into host cell to initiate an infection. The occluded baculovirus virions may cause midgut infections however on penetration into the host cells they shed this matrix, a process which facilitates cell to [url=http://www.thehygienerevolution.com/barbour.php]barbour[/url] cell infections (George, 2011). A number of early experiments has linked cell pathology caused by the viral infections to cell cycle interruption especially at the stage of cell replication where the cell cycle of the host cell is [url=http://www.achbanker.com/home.php]hollister[/url] arrested. Despite this arrest in cell multiplication the viral genome continues its multiplication [url=http://www.vivid-host.com/barbour.htm]barbour uk outlet[/url] thus raising a query on dependency of host cell mitosis in viral multiplication. For instance baculovirus genome has been utilised in development of Oryctes baculovirus which is a viral biocide used for the control of Oryctes rhinoceros insect pests.
It is necessary for characterisation of baculovirus genetic material to understand its effects to an host cell SF9 and this will boost development of an host target viral biocide of the insect pest such as Oryctes rhinoceros. As the objective of this study is to investigate theeffect of baculovirus introduction [url=http://www.gotprintsigns.com/monclerpascher/]moncler pas cher[/url] into the cell cycle of Spodoptera frugiperda (Sf9) host cells, we aim at checking cellular physiology in the sf9 cell phase distribution which is even in suspension cultures (Mohan and Gobinathan, 1992).
That is about 29% of the cells arein G1, 33% in S, and 36% [url=http://www.1855sacramento.com/woolrich.php]woolrich outlet[/url] in G2/M phase, with the duration of both G1and S phases being in an approximity of 6 hours while the duration of G2/M phase combined being about 8 hours. On Sf9 cells infection with AcMNPV, approximately 84% of exposed hostcellswere arrested in G2/M phase by 18-24 hours after viral introduction.
Results
The results obtained shows that for both the positive control and transfected cells there is signs of infection and this is characterised by cell pathological changes, such as granulation, vacuolation, polyhedron and cell enlargement. On the other hand the negative control where no introduction of viral genome was done does not show any sign of cell Pathology (CPE). Even after the arrest of [url=http://www.1855sacramento.com/peuterey.php]peuterey[/url] the host cells at the G2/M phase, the viral DNA continued to undergo replication as indicated by the CPE on the sf9 cells. The progeny virus produces unique intranuclear microvesicles on host cells, and it appears that this arrest at G2/M phase of mitosis is significant for the optimal maturation and assembly of virus
As the phosphate skeleton of DNA structure is negatively charged under neutral PH, when introduced in an electrical potential it is attracted to positive terminal, and its molecular size determines the rate of movement in the agarose gel. This is the principle used in electrophoresis and large particles show a slower rate of motion as they manipulates through agar [url=http://www.gotprintsigns.com/hollisterpascher/‎]hollister paris[/url] gel network in comparison with smaller ones. The assays of extracellular virions in insect cell culture supernatants on the third day of electrophoresis may be a significant monitor for transfection.
Discussion
This indicates that introduction of viral genome influences arrest of host cell undergoing mitosis at S-phase, leading to abnormally large nuclei, and other CPE, through alteration of cellular control of DNA replication. From the results the virus do not stimulate a "unscheduled" cellular DNA replication, then extra genes might be responsible for regulation of both baculovirus and cellular DNA replication. At the arrest phase G2/M of SF9 cells, induced by pathogen, there is concomitant detection of high [url=http://www.davidhabchy.com]barbour outlet[/url] levels of both cdc2-associated histone H1 kinase activity and cyclin B protein.
A day after infection cyclin B disappeared, but cdc2-associated histone H1 kinase activity occurred in the whole process (Mann and King, 1989). Thus prolonged regulation of cell motosis is due to protein(s) encoded by AcMNPV after the initial temporal cyclin B/cdc2 regulation.
DNA hybridisation analysis shows that maximum Viral DNA replication occurs before G2/M arrest phase, when cellular DNA replication is active. But continued replication of viral DNA which occurs several hours after the arrest indicates that, the baculovirus genome does not depend on host cell DNA replication to multiply. This biphasic nature of AcMNPV infection is organized for maximum viral DNA replication and boculovirus maturation early on infection, however on later stages of infection the process of gene regulation is directed for optimal maturation of the occluded form of viral genome.
Viral DNA replication also was not significantly altered by the introduced inhibitor fdUrd treatment, and they continue to cause CPE on the sf9 cells. It can be concluded that the infection and transfection of host cells (SF9) with a viral genome leads to development of an infectious baculovirus progeny, as demonstrated by CPE (Volkmanet al, 1976).
Conclusion
Both the positive control and the transfected host cells in the study shows CPE of vacuolation, polyhedron, cell enlargement and granulation. The changes are not seen in the negative control, where the viral genome is not introduced into the host cells. These investigation results indicates that on transfection of boculovirus DNA material into the SF9 host cells, there is an occurrence of an active infection of these cells as demonstrated by the cell pathology (Mann and King, 1989). This gives a conclusion that transfection with baculovirus genetic material leads to development of a subsequent infectious baculovirus progeny in SF9 cells. This is very important as the experiment can be used in the production of biological pesticides which are very health in the modern organic [url=http://www.sandvikfw.net/shopuk.php]hollister sale[/url] farming. As cloning [url=http://www.rtnagel.com/airjordan.php]jordan pas cher[/url] can be carried out to produce a progeny of baculovirus which targets certain type of insects thus secure and health control of pesticides can be achieved unlike the harmful use of chemical pesticides in the society.
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