Fluorescent Transgenic Silkworm
ZHANG Feng, ZHAO Yun, LU Chang-De1*
( Shanghai Institute of Biochemistry, the Chinese Academy of Seiences, Shanghai
200031, China )
CHEN Xiu, XU An-Ying, HUANG Jun-Ting
( The Sericultural Institute, Chinese Academy of Agricultural Sciences, Zhenjiang
212018, China )
Abstract The replacement of fibroin
heavy chain gene in silkworm by site directed homologous recombination was
studied. The DNA fragment, consisting of an IE promoter driving green
fluorescent protein (GFP) gene as reporter flanked by pieces of the 5′ and 3′
sequences of the fibroin heavy chain gene of silkworm at two sides, was
transferred into silkworm eggs by electroporation. Green fluorescent flecks
were seen on three silkworms fifth instar among five thousand silkworms under
UV light. PCR analysis proved that the GFP gene was integrated into the genome
of silkworm．Southern hybridization of genomic DNA of
one transgenic silkworm showed that the fibroin heavy chain gene was
successfully knocked out and replaced by the reporter gene. The transgenic
silkworms could grow up to the fifth instar as normal, but they could not spin
silk while control ones do.
Key Words GFP; fibroin heavy chain gene;
knockout
Corresponding author:
Tel, 86-21-64374430-234; Fax, 86-21-64338357