Molecular analysis of thecDNA and gene for a major protein from flexible cuticles of the giant silkmoth Hyalophora cecropia

HCCP12 is a major protein found in flexible cuticles of all three metamorphic stages of the giant silkmoth Hyalophora cecropia. The distribution of HCCP12 provides an opportunity to study the differential regulation of gene expression since there are persistent populations of epidermal cells that secrete the cuticular proteins during metamorphosis. An oligonucleotide probe, designed from N-terminal protein sequence, was used to screen a cDNA library made using mRNA of larval abdominal epidermal cells. A cDNA clone (cDNA1) was shown to encode HCCP12 by sequencing and Northern analysis. cDNA1 was used to screen a H. cecropia genomic library, and the clone II-5 was shown to contain the gene for HCCP12 by sequencing of the subclone pT7-2.8HE. This subclone contained the only fragment of II-5 that hybridized to cDNA1, indicating that HCCP12 is the only related cuticular protein gene in the 17 Kb genomic region. A set of ten GTCT repeats was found in the 5$\sp\prime$ sequence flanking the coding region for HCCP12. Searches of the GenBank database revealed that a number of other sequences contain similar repeat regions, but none of the repeats have been shown to have any functional significance. Sequence analysis by RACE (rapid amplification of cDNA ends) of the 5$\sp\prime$ ends of mRNAs for HCCP12 isolated from larval, pupal, and pharate adult epidermal cells was used to determine the transcriptional start site, and showed that cDNA 1 is ten bases short of full length. Furthermore, it was concluded that a single gene and a single promoter are used in the three metamorphic stages since the 5$\sp\prime$ ends of the mRNAs from the various tissues were identical. Thus, metamorphosis does not require stage-specific gene sets or promoters.

Name thesis:degree_name

Ph.D.

Level thesis:degree_level

Dissertation

Discipline thesis:degree_discipline

Entomology

Grantor

University of Illinois at Urbana-Champaign

Year dc:date

2011

Author dc:creator

• Binger, Lynetta C.

Contributors dc:contributor

• Willis, Judith H.

• Biology, Entomology
• Biology, Genetics
• Biology, Cell

Statement dc:rights

• Copyright 1991 Binger, Lynetta C.

Language dc:language

eng

Identifier

AAI9210746
(UMI)AAI9210746

OAI identifier oai:identifier

oai:www.ideals.illinois.edu:2142/22418