Cloning of LPAR2 Variant (ChEST973J21) for Functional Analysis

School Name

Governor's School for Science & Mathematics

Grade Level

12th Grade

Presentation Topic

Cell and Molecular Biology

Presentation Type

Mentored

Mentor

Mentor: Eric Birgbauer, Winthrop University

Abstract

Lysophosphatidic Acid (LPA) is a phospholipid derivative that is involved in several biological activities, including acting as a signaling molecule. LPA has been previously observed to cause specific, dose-dependent cone collapse of retinal neurons in chick embryonic retinal axons. LPA is expressed in various cell and tissue types. Several studies also tie LPA to human cancer, various diseases, disorders, and infertility. The LPA receptor LPA2 is encoded by the LPAR2 gene, which is expressed in several organs including the prostate, spleen, pancreas, and ovary. Recently, a chicken cDNA clone, ChEST973j21 (ChEST), was observed to be partially identical to LPAR2 at the nucleotide level, so it was considered an LPAR2 variant. It is important to understand the biological functions of ChEST in relation to cellular signaling and response. In order to do this, ChEST and chicken LPAR2 were cloned into mammalian expression vectors so they could ultimately be expressed in B103 cells to assess ChEST function. By determining the functions of ChEST, it can be compared to the cellular responses and interactions of chicken LPAR2. This will help determine whether ChEST is a new LPA receptor that responds to LPA. In order to determine this, successful ligation must first be obtained. Therefore, it is important to understand how to achieve successful ligation. Several modifications have been made to the protocol to try and obtain successful ligation. However, ligation was never successful, but amplification of the ChEST gene and expression vector were.

Location

Wall 209

Start Date

3-25-2017 9:30 AM

Presentation Format

Oral and Written

Group Project

No

COinS
 
Mar 25th, 9:30 AM

Cloning of LPAR2 Variant (ChEST973J21) for Functional Analysis

Wall 209

Lysophosphatidic Acid (LPA) is a phospholipid derivative that is involved in several biological activities, including acting as a signaling molecule. LPA has been previously observed to cause specific, dose-dependent cone collapse of retinal neurons in chick embryonic retinal axons. LPA is expressed in various cell and tissue types. Several studies also tie LPA to human cancer, various diseases, disorders, and infertility. The LPA receptor LPA2 is encoded by the LPAR2 gene, which is expressed in several organs including the prostate, spleen, pancreas, and ovary. Recently, a chicken cDNA clone, ChEST973j21 (ChEST), was observed to be partially identical to LPAR2 at the nucleotide level, so it was considered an LPAR2 variant. It is important to understand the biological functions of ChEST in relation to cellular signaling and response. In order to do this, ChEST and chicken LPAR2 were cloned into mammalian expression vectors so they could ultimately be expressed in B103 cells to assess ChEST function. By determining the functions of ChEST, it can be compared to the cellular responses and interactions of chicken LPAR2. This will help determine whether ChEST is a new LPA receptor that responds to LPA. In order to determine this, successful ligation must first be obtained. Therefore, it is important to understand how to achieve successful ligation. Several modifications have been made to the protocol to try and obtain successful ligation. However, ligation was never successful, but amplification of the ChEST gene and expression vector were.