Document Type

Open Access Thesis

Embargo Period

12-9-2016

Degree Program

Animal Science

Degree Type

Master of Science (M.S.)

Year Degree Awarded

2017

Month Degree Awarded

February

Advisor Name

Cynthia

Advisor Middle Initial

L

Advisor Last Name

Baldwin

Abstract

Workshop cluster 1 (WC1) molecules are exclusively expressed on the surface of gamma delta T cells and act as co-receptors and bind pathogens thus also functioning as pattern recognition receptors. The aim was to obtain cDNA evidence to support the recent caprine genome annotation of the WC1 multigene family conducted by a colleague. To get cDNA sequences three strategies were used. Strategy 'I' was used to obtain three clones that corresponded to WC1 SRCR domain d9 through the intracytoplasmic tail sequence. Strategy 'II' was used to obtain 6 clones. A PCR was conducted using SRCR domain b7 through the intracytoplasmic tail sequence. A third strategy obtained full-length WC1 transcripts. The three sequences that extended from SRCR domain d9 to the intracytoplasmic tail matched closely with predicted goat Gene 1 or 14. Another 3 sequences that extended from the SRCR b7 domain through SRCR domain d11 or through the intracytoplasmic tail matched with the predicted Genes 1, 2 and 14, respectively. Two additional full-length cDNA clones CH-MA-03 and 41 were completely sequenced in stages which involved a PCR amplication of the internal domains to complete the sequencing. The a1 domain of CH-MA41 was 100% similar to the annotated and predicted Gene 4 while CH-MA03 also was closest to Gene 4 with a 99% similarity. However, the intracytoplasmic tail sequence of these two cDNA clones was a Type II tail while Gene 4 had a Type I tail. Because of this difference in tails these two cDNA clones had a greater overall similarity with Genes 7 and 15 which had Type II tails. These results suggest that the genome assembly may have errors.

First Advisor

Cynthia L. Baldwin

Share

COinS