Trehalose Plays Role in cDNA Research

by J. C. Spencer

One word that is continually attributed to the sugar trehalose is STABLE. It helps stabilize cells, enzymes, proteins, DNA and the research goes on. Trehalose is used in synthesizing full length cDNA because it helps enzymes maintain their activity at higher temperatures where they normally become inactive.

Here is the research carried out in Japan.

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Biochemistry

Thermostabilization and thermoactivation of thermolabile enzymes by trehalose and its application for the synthesis of full length cDNA

Piero Carninci,* Yoko Nishiyama,* Arthur Westover,* Masayoshi Itoh,* Sumiharu Nagaoka,* Nobuya Sasaki,* Yasushi Okazaki,* Masami Muramatsu,* and Yoshihide Hayashizaki*†‡

*Genome Science Laboratory, Tsukuba Life Science Centre, The Institute of Physical and Chemical Research (RIKEN), 3-1-1 Koyadai, Tsukuba, Ibaraki 305, Japan; and †Tsukuba University Medical School, 1-1-1, Tennodai, Tsukuba, Ibaraki 305, Japan

‡To whom reprint requests should be addressed.

Communicated by Takashi Sugimura, National Cancer Center, Tokyo, Japan

Abstract:
The advent of thermostable enzymes has led to great advances in molecular biology, such as the development of PCR and ligase chain reaction. However, isolation of naturally thermostable enzymes has been restricted to those existing in thermophylic bacteria. Here, we show that the disaccharide trehalose enables enzymes to maintain their normal activity (thermostabilization) or even to increase activity at high temperatures (thermoactivation) at which they are normally inactive. We also demonstrate how enzyme thermoactivation can improve the reverse transcriptase reaction. In fact, thermoactivated reverse transcriptase, which displays full activity even at 60°C, was powerful enough to synthesize full length cDNA without the early termination usually induced by stable secondary structures of mRNA.

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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC18452/

View the cDNA animation
http://www.maxanim.com/genetics/cDNA/cDNA.htm

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