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Volume 271, Number 45, Issue of November 8, 1996 pp. 28017-28023
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.

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Organization of Serpin Gene-1 from Manduca sexta
EVOLUTION OF A FAMILY OF ALTERNATE EXONS ENCODING THE REACTIVE SITE LOOP

(Received for publication, May 28, 1996, and in revised form, August 30, 1996)

Haobo Jiang , Yang Wang , Yanling Huang , Amy B. Mulnix ^¶ , Jeannette Kadel , Kenneth Cole and Michael R. Kanost

From the  Department of Biochemistry, Kansas State University, Manhattan, Kansas 66506, the ^¶ Department of Biology, Earlham College, Richmond, Indiana 47374, and the  Biotechnology Division, National Institute of Standards and Technology, Gaithersburg, Maryland 20899

Manduca sexta serpin gene-1 encodes a family of serpins whose amino acid sequences are identical in their amino-terminal 336 residues but variable in their carboxyl-terminal 39-46 residues, which includes the reactive site loop (Jiang, H., Wang, Y., and Kanost, M. R. (1994) J. Biol. Chem. 269, 55-58). Here, we report the gene's complete nucleotide sequence and exon-intron structure. A unique characteristic of this gene is its exon 9, which is present in 12 alternate forms between exons 8 and 10. Isolation and characterization of cDNA clones containing exons 9C, 9H, and 9I, which were not found previously, indicate that all 12 alternate forms of exon 9 can be utilized to generate 12 different serpins. The splicing pathway apparently allows inclusion of only one exon 9 per molecule of mature serpin-1 mRNA. Analysis of exon-intron border sequences reveals unique features that may be involved in regulation of RNA splicing. The exon 9 region has apparently evolved through rounds of exon duplication and sequence divergence. The exons near the center of the region may have evolved recently, whereas the outermost exons are the most ancient. Exons 9G and 9H were duplicated as a pair from exons 9E and 9F, an event that may have occurred more than once in the history of this gene.

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