phiC31-integrase-mediated, site-specific integration of transgenes in the silkworm, Bombyx mori (Lepidoptera: Bombycidae)

Naoyuki Yonemura, Toshiki Tamura, Keiro Uchino, Isao Kobayashi, Ken ichiro Tatematsu, Tetsuya Iizuka, Takuya Tsubota, Hideki Sezutsu, Mayandi Muthulakshmi, Javaregowda Nagaraju, Takahiro Kusakabe

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Transgenic silkworms can be useful for investigating the functions of genes in the post-genomic era. However, the common method of using a transposon as an insertion tool may result in the random integration of a foreign gene into the genome and suffer from a strong position effect. To overcome these problems, it is necessary to develop a site-specific integration system. It is known that phiC31 integrase has the capacity to mediate recombination between the target sequences attP and attB. To test the availability of site-specific integration in the silkworm, we first examined the efficiency of recombination between the target sites of the two plasmids in silkworm embryos and found that the frequency of recombination was very high. Then we constructed a host strain that possessed the target sequence attP using the common method. We injected the donor plasmid together with the phiC31 integrase mRNA into the embryos of the host strain and obtained positive lines. Structural analysis of the lines showed that site-specific integration occurred by recombination between the genomic attP site and the attB site of the donor plasmid. We can conclude from the results that phiC31 integrase has the ability to mediate the site-specific integration of transgenes into the silkworm chromosome.

Original languageEnglish
Pages (from-to)265-273
Number of pages9
JournalApplied Entomology and Zoology
Volume48
Issue number3
DOIs
Publication statusPublished - Aug 1 2013

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Bombycidae
Bombyx mori
silkworms
transgenes
Lepidoptera
host strains
plasmids
position effect (genetics)
genomics
transposons
genes
genetically modified organisms
chromosomes
genome
methodology
testing

All Science Journal Classification (ASJC) codes

  • Insect Science

Cite this

phiC31-integrase-mediated, site-specific integration of transgenes in the silkworm, Bombyx mori (Lepidoptera : Bombycidae). / Yonemura, Naoyuki; Tamura, Toshiki; Uchino, Keiro; Kobayashi, Isao; Tatematsu, Ken ichiro; Iizuka, Tetsuya; Tsubota, Takuya; Sezutsu, Hideki; Muthulakshmi, Mayandi; Nagaraju, Javaregowda; Kusakabe, Takahiro.

In: Applied Entomology and Zoology, Vol. 48, No. 3, 01.08.2013, p. 265-273.

Research output: Contribution to journalArticle

Yonemura, N, Tamura, T, Uchino, K, Kobayashi, I, Tatematsu, KI, Iizuka, T, Tsubota, T, Sezutsu, H, Muthulakshmi, M, Nagaraju, J & Kusakabe, T 2013, 'phiC31-integrase-mediated, site-specific integration of transgenes in the silkworm, Bombyx mori (Lepidoptera: Bombycidae)', Applied Entomology and Zoology, vol. 48, no. 3, pp. 265-273. https://doi.org/10.1007/s13355-013-0182-6
Yonemura, Naoyuki ; Tamura, Toshiki ; Uchino, Keiro ; Kobayashi, Isao ; Tatematsu, Ken ichiro ; Iizuka, Tetsuya ; Tsubota, Takuya ; Sezutsu, Hideki ; Muthulakshmi, Mayandi ; Nagaraju, Javaregowda ; Kusakabe, Takahiro. / phiC31-integrase-mediated, site-specific integration of transgenes in the silkworm, Bombyx mori (Lepidoptera : Bombycidae). In: Applied Entomology and Zoology. 2013 ; Vol. 48, No. 3. pp. 265-273.
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