Enzyme Expression

1. Enzyme Expression Enzyme Expression Creative Enzymes provides custom services for expression and production of both natural and recombinant enzymes in various systems. As understanding of enzymology and molecular biology progresses, enzymes have been found more and more important in almost all biological pathways. At this point, there are over 200 protein-based pharmaceuticals licensed by FDA. Sales of therapeutic enzymes and industrial enzymes summed to over $6 billion by 2010. Rapid growth in the market demands and product development requires services of large-scale production. Meanwhile, new enzymes are being developed to meet all types of needs in pharmaceutical and catalysis. Advanced expression of enzymes with special properties and functions is desired by all researchers. Creative Enzymes established and developed expression systems for all types of enzymes, covering early stage of cDNA preparation, DNA cloning, enzyme expression, all the way to production scale-up. Our outstanding scientists and strategic partners professionally handle every detail of each step. We are the experts you can trust in the whole process of enzyme expression. Several steps are necessary to obtain a natural or recombinant enzyme depending on the source of the gene sequence and the specific technical requirement: The initial cDNA synthesis, amplification, and isolation; followed by insertion into a vector; and lastly induction and expression of the target enzyme (Figure 1). The first two steps are also usually considered as cDNA cloning. A desired enzyme could be a natural enzyme existing in high abundance in specific cells at a certain time, which is generally related to high abundance of the corresponding mRNA. Therefore, the mRNA can be identified and isolated to be used as the template for reverse transcription to generate cDNA, which is complementary to the mRNA sequence and does not contain introns or operons. Alternatively, if the coding sequence of the enzyme is known, fully synthesize cDNA sometimes is a fast and more economic approach. After obtaining the target cDNA, amplification of the cDNA copies is carried out. Traditionally, a cDNA copy is inserted into a vector and host cells are used to multiply the vector DNA, also known as recombinant DNA, along with the DNA of the host cells. The subsequent isolation and purification will give the target cDNA in a large quantity. However, multiple steps of cutting and ligation, as well as purification of DNA are involved in this process, resulting in high cost and low efficiency. Thus, the PCR (polymerase chain reaction) technology has revolutionized cDNA cloning since it was invented. The technology makes DNA cloning much faster and easier for both particular DNA clones and construction of a cDNA library. With recent development of the technology, such as qPCR

2. (quantitative PCR) and RT-PCR (reverse transcription PCR), it has become the most commonly used and often indispensable tool in cDNA cloning.