Title of Oral/Poster Presentation

Isolating a Gene for Spider glue

Class

Article

Department

Biology

Faculty Mentor

Randolph Lewis

Presentation Type

Poster Presentation

Abstract

The spider aggregate gland from orb weaving spiders produces a glue that helps bind insects to the web. Very little is known about the glue, but possible applications for a synthetically produced aggregate protein range from underwater adhesives to surgical glue. We are cloning the aggregate cDNA to confirm and extend existing knowledge about the aggregate gene(s). mRNA was extracted from aggregate glands using TRIzol reagent, and then reverse transcribed using MMLV-RT to create single stranded DNA. The second strand was next synthesized employing DNA Polymerase I in conjunction with RNaseH to form cDNA. The cDNA was ligated into pBluescript II and transformed into electrocompetent DH10B bacterial cells. Bacteria were plated on ampicillin and plates used had a colony density of approximately 1000 colonies per plate. Libraries screening has commenced with a probe designed using data from two previously reported sequences for the aggregate gene. Preliminary screening of the library was unsuccessful, but the second highly efficient library is now available for further screening. We expect to obtain 2.5-3Kb of clear aggregate sequence from this library in order to confirm previous sequences and allow us to express this gene synthetically.

Start Date

4-9-2015 3:00 PM

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Apr 9th, 3:00 PM

Isolating a Gene for Spider glue

The spider aggregate gland from orb weaving spiders produces a glue that helps bind insects to the web. Very little is known about the glue, but possible applications for a synthetically produced aggregate protein range from underwater adhesives to surgical glue. We are cloning the aggregate cDNA to confirm and extend existing knowledge about the aggregate gene(s). mRNA was extracted from aggregate glands using TRIzol reagent, and then reverse transcribed using MMLV-RT to create single stranded DNA. The second strand was next synthesized employing DNA Polymerase I in conjunction with RNaseH to form cDNA. The cDNA was ligated into pBluescript II and transformed into electrocompetent DH10B bacterial cells. Bacteria were plated on ampicillin and plates used had a colony density of approximately 1000 colonies per plate. Libraries screening has commenced with a probe designed using data from two previously reported sequences for the aggregate gene. Preliminary screening of the library was unsuccessful, but the second highly efficient library is now available for further screening. We expect to obtain 2.5-3Kb of clear aggregate sequence from this library in order to confirm previous sequences and allow us to express this gene synthetically.