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11. Molecular Cloning and Analysis of theTryptophan oxygenaseGene in the Silkworm,Bombyx mori

Author

Shen WeiFeng, Meng ZhiQi, Weng Hongbiao, Niu Baolong, Liu Yan, and He Lihua

Subjects
Molecular Sequence Data, Molecular cloning, Gene Expression Regulation, Enzymologic, Protein Structure, Secondary, Article, Bombycidae, eye pigmentation, vermilion gene, Bombyx mori, Complementary DNA, Animals, Amino Acid Sequence, Cloning, Molecular, Peptide sequence, Gene, Phylogeny, Base Sequence, Molecular mass, biology, Gene Expression Profiling, fungi, General Medicine, Bombyx, biology.organism_classification, Eye pigmentation, Molecular biology, Tryptophan Oxygenase, Lepidoptera, Biochemistry, Insect Science, Sequence Alignment
Abstract

A Bombyx mori L. (Lepidoptera: Bombycidae) gene encoding tryptophan oxygenase has been molecularly cloned and analyzed. The tryptophan oxygenase cDNA had 1374 nucleotides that encoded a 401 amino acid protein with an estimated molecular mass of 46.47 kDa and a PI of 5.88. RT-PCR analysis showed that the B. mori tryptophan oxygenase gene was transcribed in all examined stages. Tryptophan oxygenase proteins are relatively well conserved among different orders of arthropods.

Published
2008

13. Germline Transformation of the Silkworm Bombyx moriL. by Sperm-Mediated Gene Transfer1

Author

Zhou, Wenlin, Gong, Chengliang, Xue, Renyu, Cao, Guangli, Cao, Jinru, Ye, Aihong, Weng, Hongbiao, and Wang, Yongqiang

Abstract

The domesticated silkworm Bombyx moriL. has important roles in basic biological research and applied science. To explore the practical use of transgenic technology in agricultural silkworm varieties, we fused the neomycin-resistance gene (NeoR) and the green fluorescent protein gene (gfp) into the piggyBac-based transposon vector and transduced it into silkworms by sperm-mediated gene transfer (SMGT). Fluorescence observation indicated the positive rate of G0 egg-batches is 72.7%. After screening against the antibiotic G418, development of individual larvae in the same brood showed significant size differences. PCR detection indicated the existence of gfpand NeoRand confirmed the positive rate of transgenesis as 0.47%. Southern blot analysis confirmed the presence of the exogenous genes in the genome of G7 larvae. These results show that our strategy is practical and markedly improves the efficiency of SMGT.

Published
2012
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