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An optimized
vip3A
gene, designated as
vip3A*
was chemically synthesized and a
thi1
gene chloroplast transit peptide coding sequence was attached to its 5′ end to produce the
tvip3A*
.
vip3A*
and
tvip3A*
genes were transformed into
Gossypium hirsutum
cv. Zhongmiansuo35 mediated by
Agrobacterium tumefaciens
. Four independent transgenic T1 lines with single-copy insertions and unchanged phenotypes (CTV1 and CTV2 for
tvip3A*
, and CV1 and CV2 for
vip3A*
) were selected by Polymerase chain reaction (
PCR
), Reverse transcription (RT)-PCR, Southern blotting, enzyme-linked immunosorbent assay (ELISA), and insect bioassay. As expected, the Vip3A* protein of CTV1 and CTV2 were transported to the chloroplasts, where they accumulated. Our results suggest that the two
tvip3A*
transgenic lines (CTV1 and CTV2) can be used to develop insect-resistant cultivars and could be used as a resource for raising multi-toxins-expressing transgenic cotton.
