Dr. Stephen Baron
Degree Award Date
Polymerase Chain Reaction, Cloning, sequencing, Polyhydroxybutyrate Depolymerase Gene, Streptomyces sp. 5A, DNA probe, southern blotting technique, Diverging non-degenerate primers, inverse PCR
Biology | Cell and Developmental Biology | Molecular Genetics
A .500 base pair (bp) segment of the polyhydroxybutyrate PHB depolymerase gene (phaZ) of Streptomyces sp. 5A was amplified by the polymerase chain reaction (PCR), cloned, and sequenced. Homology searches suggested that this segment (A20TG) comprised about 40% of the length of phaZ. Specific primers for A20TG were then used in PCR to construct a phaZ-specific DNA probe. The probe was used in the southern blotting technique in order to identify restriction fragments suitable for cloning the remainder of phaZ and its upstream region; however, results from this method could not be analyzed and interpreted with certainty. Diverging non-degenerate primers complementary to A20TG were used in an alternative method, inverse PCR, in attempts to clone the remainder of the gene. We have obtained PCR products which will be cloned and sequenced for this purpose.
Griffin, Tessa A., "Use of the Polymerase Chain Reaction to Clone Sections of the Polyhydroxybutyrate Depolymerase Gene of Streptomyces sp. 5A" (2007). Honors Projects. 187.