[Construction of the synthetic genes for protein analogs of spider silk carcass spidroin 1 and their expression in tobacco plants].
To obtain transgenic tobacco plants expressing recombinant analogs of spider dragline silk spidroin 1, artificial 1f5 and 1f9 coding for spidroin 1 analogs were 3'-fused in-frame with the reporter lichenase gene. The Tr2' weak constitutive promoter of Agrobacterium tumefaciens T-DNA and the strong constitutive promoter of the cauliflower mosaic virus 35S RNA gene were used as regulatory elements. The expression cassettes were used to transform agrobacteria and then introduced in tobacco leaf disks. On evidence of Southern hybridization, transgenic plants each carried a single copy of a hybrid gene, which corresponded in size to the constructed one. Zymography and Western blotting revealed full-length hybrid proteins in leaf extracts of transgenic plants. The results testified that plants can maintain and express synthetic genes for spider silks and, consequently, may be used as a convenient producer of recombinant silk analogs.
Piruzian
E S
ES
Vavilov Institute of General Genetics, Russian Academy of Sciences, Moscow, 119991 Russia. eleopiru@vigg.ru
Bogush
V G
VG
Sidoruk
K V
KV
Goldenkova
I V
IV
Musiĭchuk
K A
KA
Debabov
V G
VG
D016960:Agrobacterium tumefaciens / Q000235:genetics; D000818:Animals; D017796:Caulimovirus / Q000235:genetics; D005349:Fibroins; D005786:Gene Expression Regulation; D005822:Genetic Vectors; D006026:Glycoside Hydrolases / Q000235:genetics; D019476:Insect Proteins / Q000737:chemistry / Q000235:genetics*; D018515:Plant Leaves / Q000235:genetics; D030821:Plants, Genetically Modified / Q000235:genetics*; D011401:Promoter Regions, Genetic; D015202:Protein Engineering / Q000379:methods*; D011506:Proteins / Q000235:genetics*; D012367:RNA, Viral; D038621:Regulatory Sequences, Ribonucleic Acid; D047011:Silk; D013112:Spiders; D014026:Nicotiana / Q000235:genetics*
D019476:Insect Proteins; D011506:Proteins; D012367:RNA, Viral; D038621:Regulatory Sequences, Ribonucleic Acid; D047011:Silk; C086087:spidroin 1; D005349:Fibroins; D006026:Glycoside Hydrolases; C028020:licheninase