STRUCTURE AND DEVELOPMENTAL EXPRESSION OF A LARVAL CUTICLE PROTEIN GENE OF THE SILKWORM, BOMBYX-MORI

被引:33
作者
NAKATO, H [1 ]
SHOFUDA, K [1 ]
IZUMI, S [1 ]
TOMINO, S [1 ]
机构
[1] TOKYO METROPOLITAN UNIV, DEPT BIOL, HACHIOJI, TOKYO 19203, JAPAN
来源
BIOCHIMICA ET BIOPHYSICA ACTA-GENE STRUCTURE AND EXPRESSION | 1994年 / 1218卷 / 01期
关键词
CUTICLE PROTEIN; GENE CLONING; NORTHERN HYBRIDIZATION; S1 NUCLEASE MAPPING; JUVENILE HORMONE; (B-MORI);
D O I
10.1016/0167-4781(94)90101-5
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Structure and expression of the gene for a larval cuticle protein of the silkworm, Bombyx mori were studied. A major cuticle protein, termed 'LCP30' was purified from the urea extract of integuments of the fifth (final) instar larvae. Immunoblot analysis by use of the anti-LCP30 antibody revealed that LCP30 begins to accumulate in larvae as early as 10 h after hatch and is present throughout the,larval stages. The LCP30 epitope is also detectable in the adult abdominal integument but is absent from pupal integument and adult wing. Screening of an epidermal cDNA expression library with the antibody probe yielded a cDNA clone for LCP30. Primary structure deduced from the cDNA sequence showed that LCP30 bears an arginine-glycine-aspartate (RGD) sequence. The region around this domain exhibits striking similarity with the amino acid sequences found in vertebrate collagens. The genomic DNA clone coding for LCP30 was isolated by screening a B. mori gene library with the LCP30 cDNA probe. The gene consists of five exons interspersed by four introns spanning over 2.7 kb region of chromosomal DNA. The LCP30 mRNA is detectable at high levels at larval intermolt stages, gradually declines after the fourth molt and totally disappears at mid-fifth larval instar, indicating that the expression of LCP30 gene is regulated in a stage-specific fashion in the epidermal cells. Topical application of a juvenile hormone analogue (methoprene) to the fifth instar larvae followed by RNA blot and S1 nuclease mapping analyses of the epidermal RNA proved that juvenile hormone activates transcription of the LCP30 gene.
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页码:64 / 74
页数:11
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