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Isolation and characterization of two cDNAs expressed in the Harderian gland of the anole lizard ( Anolis carolinensis ) (589.4)
Author(s) -
Prokopiak Zoey,
Steglich Carolyn
Publication year - 2014
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.28.1_supplement.589.4
Subject(s) - biology , complementary dna , anolis , gene , harderian gland , lizard , genome , genbank , microbiology and biotechnology , genetics , zoology , endocrinology
The Harderian gland (HG) is a secretory gland in the eye of most land animals. Its structure has been studied, but little is known about the gland's actual function, which varies depending on species. Structural observations have suggested multiple possible functions (lubrication of the eye, sensing of chemical signals via the vomeronasal organ, or an immunological role) but have little supportive experimental data. To investigate the HG’s function in an organism such as the Anole lizard (Anolis carolinensis), cDNAs were isolated from the HG and sequenced. Two cDNAs (E1 and M1) that code for proteins of unknown structure or function (no significant hits in a GenBank search) were selected for further study. Using RT‐qPCR (reverse transcriptase quantitative polymerase chain reaction), the expression of the cloned genes (cDNAs) that had been isolated from the HG of the Anole lizard were compared to other known Anolis genes (in this case, genes encoding ribosomal proteins) that are expressed in all cells. The E1 cDNA is expressed at high levels in the HG but also expressed at background levels in liver and tongue tissues. The M1 cDNA is expressed at extremely high levels only in the HG. These cDNAs have been subcloned into the pET‐22 expression vector for expression in bacterial cells to provide an unlimited source of the proteins. The functional characterization of the proteins can then be directly investigated. The genome of the Anole lizard was completed in 2010 but is currently incompletely annotated. BLAST searches of the Anole's genome with the E1 and M1 sequences have returned some short matches for the sequences, which locates each of them to a particular scaffold in the genome, but no predicted genes are annotated for those regions. Additional alignment work with MegAlign locates the likely exons and introns of these genes more precisely, thereby providing additional annotation to those regions of the Anole genome. Grant Funding Source : Supported by a Faculty/Student Research Grant and a Student Research Grant from Slippery Rock Univ.