• Title/Summary/Keyword: Cloning and Expression

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Cloning and mRNA Expression of an Actin cDNA from the Mulberry Longicorn Beetle, Apriona germari

  • Gui, Zhongzheng;Lee, Kwang Sik;Wei, Yadong;Yoon, Hyung Joo;Kim, Iksoo;Guo, Xijie;Sohn, Hung Dae;Jin, Byung Rae
    • International Journal of Industrial Entomology and Biomaterials
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    • v.9 no.2
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    • pp.187-191
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    • 2004

  • Actin is a ubiquitous and highly conserved protein found in eukaryotic organisms. In this study, we describe the cDNA cloning and mRNA expression of an actin gene from the mulberry longicorn beetle, Apriona germari. The A. germari actin cDNA is 1524 bp containing a complete 1128 bp open reading frame that encodes a polypeptide of 376 amino acid residues with a predicted molecular weight of about 41.5 kDa. The deduced amino acid sequence of the A.germari actin cDNA showed 99% protein sequence identity to Homalodisca coagulata actin, differing at only two amino acid positions, and 92-98% protein sequence identity to known insect species actins. The predicted three-dimensional structure of A. germari actin revealed the four residue hydrophobic pulg loop characteristic of the actin family. Northern blot analysis showed that A. germari actin is highly expressed in epidermis and muscle, and less strongly in midgut, but not in the fat body of A. germari larva.

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Molecular Cloning and Expression of a Xylanase Gene from Alkalophilic Bacillus sp.

  • Yu, Ju-Hyun;Kang, Yun-Sook;Park, Young-Seo;Bai, Dong-Hoon
    • Journal of Microbiology and Biotechnology
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    • v.1 no.4
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    • pp.251-255
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    • 1991

  • A 16 kilobase (kb) HindIII fragment of alkalophilic Bacillus sp. YC-335 containing a gene for xylanase synthesis was inserted at the HindIII site of pBR322 and cloned in Escherichia coli HB101. After subcloning of recombinant plasmid pYS52, the 1.5 kb fragment was found to code for xylanase activity, and the hybrid plasmid was named pYS55. The DNA insert of the plasmid was subjected to restriction enzyme mapping, which showed that pYS55 had single site for PuvII and SstI in the 1.5 kb insert fragment. Southern hybridization analysis revealed that the cloned gene was hybridized with chromosomal DNA from alkalophilic Bacillus sp. YC-335. About 64% of the enzyme activity was observed in the extracellular and periplasmic space of E. coli HB10l carrying pYS55.

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