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Lentivirus-mediated generation of mosaic transgenic fish
F. M. P. Tonelli, S. M. S. N. Lacerda, B. L. S. Lemos, G. M. J. Costa, L. R. Fran?a, R. R. Resende
Fish transgenesis is performed since the beginning of the 80’s by using the standard technique: microinjection. Commonly, a mosaic animal is first generated and then full transgenic organisms (fishes). However, microinjection is a technique that possesses low efficiency rate and requires highly trained people to inject correctly using the glass needle and the micromanipulator. So, in the present study we genetically modified Nile tilapias using lentiviral particles, in order to obtain a protocol to generate the mosaic transgenic animal in a faster and easier way. The lentivirus were generated in 293FT cells by using a plasmid DNA developed from pLenti6.3/V5(TM)-TOPO(R) to contain the DsRed2 fluorescent protein codifying sequence. Nile tilapia newly fertilized eggs were exposed to different concentrations of lentivirus particles (from 6.0x105 TU/mL to 6.0x104 TU/mL) for 24 hours (under agitation at 28?C) in two different systems, using Petri dishes or Falcon tubes. After that, the eggs were kept under agitation in tank water until the hatching. Larvae were analyzed under fluorescence microscope during its development and the presence of DsRed2 fluorescent protein could be noticed. The death rate was reduced when the Petri dish system was used. The protein presence was also accessed through immunohistochemistry using the antibodies Living Colors Anti-RCFP Polyclonal Pan Antibody (primary 1:1000) and Goat anti-Rabbit IgG HRP conjugate (secondary 1:500). The desired codifying sequence presence in the mosaic fishes was confirmed by analyses of mRNA through RT-PCR. So, lentivirus particles are important tools for the development of new biotechnologies in aquaculture as they can be used to quickly generate mosaic transgenic tilapias: which can be later outcrossed to produce the full transgenic fish, or which can provide gametes containing the transgene to obtain the full transgenic animal through in vitro fertilization.
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Animal Reproduction Colégio Brasileiro de Reprodu??o Animal - CBRAAv. Cel. José Dias Bicalho 1224, Lj4CEP:
- Belo Horizonte, MGFone: (+55 31)
animreprod@cbra.org.brNotifications
The Effect of Financial Crisis and Macroeconomic Factors on FDI in Developing Countries
Anita Hasli, Nurhani Aba Ibrahim, Catherine S.F. Ho
Developing countries needs FDI to be at par with the progress of developed countries. The dearth of study on the effects of financial crisis on FDI justifies the objective which is to examine the potential effect of financial crisis inclusive of macroeconomic factors as control variables on FDI in dataset 23 developing countries for the period . This study includes descriptive analysis, correlation test, stationary test and regression analysis. The random effects generalized least square (GLS) estimator is used in the regression to examine the potential effect of financial crisis and macroeconomic factors on the inflow of FDI. Foremost, the US financial crisis has a positive significance to the inflow of FDI which validates Krugman’s theory on fire-sale FDI. However, country specific economic recession, lending rates and natural resources discourage inflow of FDI. Nonetheless, trade openness, domestic currency, money supply and domestic fixed investment encourage FDI in developing countries.Keywords: Foreign Direct I REM; developing countriesJEL Classifications: C33; F21; G01; P52
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