Construction and screening of A cDNA library for the C3 gene(s) of the Nurse Shark (Ginglymostoma Cirratum)

Authors

Janette Cristina Builes, Florida International University

Document Type

Thesis

Degree

Master of Science (MS)

Major/Program

Please see currently inactive department below.

Major/Program

Medical Laboratory Science

First Advisor's Name

Sylvia L. Smith

First Advisor's Committee Title

Committee Chair

Second Advisor's Name

Charles H. Bigger

Third Advisor's Name

Janet A. Lineback

Date of Defense

11-28-2001

Abstract

Mammalian C3 is a complement protein which consists of an α chain (125kDa) and β chain (75kDa) held together by a disulfide bond. The a chain contains a conserved thiolester site which provides the molecule with opsonic properties. The protein is synthesized as a single pro-C3 molecule which is post-translationally modified. C3 genes have been identified in organisms from different phyla, however, the shark C3 gene remains to be cloned. Sequence data from the shark will contribute to understanding further the evolution of this key protein. To obtain additional sequence data for shark C3 genes a cDNA library was constructed and screened with a DIG-labeled C3 probe. Fifty clones were isolated and sequenced. Analysis identified four sequences that yielded positive alignments with C3 of a variety of organisms including human C3. Deduced amino acid sequence analysis confirmed a β/α cut site (RRRR), the CR3 and properdin binding sites, the catalytic histidine, and the reactive thiolester sequence. In the shark there are at least two C3-like genes as the gene sequence obtained is distinct from that previously described.

Identifier

FI14051873

Recommended Citation

Builes, Janette Cristina, "Construction and screening of A cDNA library for the C3 gene(s) of the Nurse Shark (Ginglymostoma Cirratum)" (2001). FIU Electronic Theses and Dissertations. 1940.
https://digitalcommons.fiu.edu/etd/1940