Increased yield of heterologous viral glycoprotein in the seeds of homozygous transgenic XXXX plants cultivated underground.

@entity1 @entity286 @entity72293 @entity4538 @entity579 @entity72292
The use of transgenic plants in the production of recombinant proteins for @entity1 therapy, including subunit vaccines, is being investigated to evaluate the efficacy and safety of these emerging biopharmaceutical products. We have previously shown that synthesis of recombinant glycoprotein B (gB) of @entity4538 can be targeted to seeds of transgenic @entity286 when directed by the @entity579 glutelin 3 promoter, with gB retaining critical features of immunological reactivity (E.S. Tackaberry et al. 1999. Vaccine, 17: 3020-3029). Here, we report development of second generation transgenic plant lines (T1) homozygous for the transgene. Twenty progeny plants from two lines ( @entity72292 (1)-2 and @entity72293 (1)-3) were grown underground in an environmentally contained mine shaft. Based on yields of gB in their seeds, the @entity72292 (1)-2 line was then selected for scale-up in the same facility. Analyses of mature seeds by ELISA showedthat gB specific activity in @entity72292 (1)-2 seeds was over 30-fold greater than the best T0 plants from the same transformation series, representing 1.07% total seed protein. These data demonstrate stable inheritance, an absence of transgene inactivation, and enhanced levels of gB expression in a homozygous second generation plant line. They also provide evidence for the suitability of using this environmentally secure facility to grow transgenic plants producing therapeutic biopharmaceuticals.
