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The total secretory proteins of the transformants were extracted, separated, and analyzed by PAGE and Western blot.

For genomic PCR, crude DNA of the WT and transformants were extracted by use of Instagene Matrix (Bio-Rad, USA) following modified protocols originally developed by Wan et al. [ 42].

Genomic DNA of the transformants was extracted as described above and digested with restriction enzymes BamHI, PstI, and NdeI (all purchased from Thermo Scientific, Rockford, IL, USA).

Molecular analysis of transformants Genomic DNA of the transformants was extracted using a phenol chloroform method (Seiboth et al. 2002).

Genomic DNA from the putative transformants was extracted from young folded leaves with DNeasy® Plant Mini Kit (Qiagen).

Plasmid DNA of the donors, transconjugants and transformants was extracted with the QIAGEN (Hilden, Germany) Plasmid Midi kit according to the manufacturer's instructions.

Genomic DNA from the transformants was extracted, digested with XhoI and NheI separately, circularized by ligation, and electroporated into E. coli XL1-Blue Electroporation-Competent Cells (Agilent Technologies) following the manufacturer's protocol.

The plasmids were extracted from E.coli transformants, purified and used to transform again the deletion mutant S. cerevisiae BY4741_ haa1∆ and also the parental strain BY4741.

Recombinant Bombyx mori nucleopolyhedrovirus (rBmNPV-NcSAG1) bacmids were extracted from white transformants and insertion of the NcSAG1 gene was confirmed by PCR.

Recombinant vectors harbouring fusion genes were extracted from E. coli transformants and purified as described above.

Recombinant vectors harbouring PCR products were extracted from E. coli transformants and purified using a Plasmid Isolation Kit (BioFlux).