| dc.description.abstract | The development of the biotechnology has permitted to achieve, among other
biotechnological products, many recombinant proteins which are largely used as vaccines,
hormones, enzymes and other ways of human therapeutic activity. This is possible because
the multidisciplinary: through the technology of the recombinant DNA the region that encode
gene of interest is cloned in a system of adequate expression for super-expression of the
correspondent protein and, through the technology of bioprocess is possible to increase the
obtaining scale of the recombinant product assuring its reproducibility, integrity and
functionality. Involving these two areas of knowledge, this works intends to express an
heterolog protein, the Canacistatina, deriving from sugar cane, through an eucariotic
expression system Sf9/baculovirus. This implies in the construction of a recombining
baculovirus, containing the cDNA which codifies to the Canacistatina, and also the
characterization of the cultivation of Sf9 cells, and of the process of infection by baculovirus,
in kinetics and physiologic therms. In order to obtain these purposes it was done experiments
of cultivation of Sf9 cells in Schott bottles of 100 mL, with work volume of 20 mL, in 27° C.
In these experiments it was analyzed the optimum conditions of velocity agitation, inoculum s
density, culture environment, and also the role of substrates. In relation to these parameters, it
was observed that a better cell growth can be conduced in a 100 rpm of agitation, in an
inoculum s density of 1x106 cell/ml, using a combination of 1:1 between the mediums SF900
II and TNM-FH (Medium 50%), without the necessity of addition of Bovine Fetal Sorum.
This last aspect represents a great advantage in relation to the bioprocesses, in a way that it
uses Sf9 cells already known. It was construed the recombining baculovírus containing the
Canacistatina s gene through the modified genome of the baculovirus Autographa californica
Multiple Nuclear Polihedrosis Virus and the infection experiments of Sf9 cells was made
under three MOI values (1, 5 e 10 pfu/cell) in the two culture mediums. The betters results
indicates to infection of Sf9 cells in 50% Medium using MOI= 5 pfu/cell, conditions that the
Canacistatina s productivity reached 28 µg/106cel. | |
