Sequence Analysis Of Protein And Its Gene Of ICL Of Suillus Luteus (L.:Fr) Gray

Isocitrate Lyase is the key enzyme of glyoxylate cycle. In microbiology, there is mechanism of isocitrate lyase:when sufficient energy reserves isocitrate lyase catalytic produce succinic acid and glyoxalic acid. When a microbe absence of glucose, the carbon dioxide will be incorporated as the sole carbon source into the glyoxylate cycle synthesized extracellular substances, so that the microorganisms survive in the hostile environment. Isocitrate lyase for Suillus luteus secrete oxalic acid, adjust pH environment, improve the phosphorus nutrition, improve the disease-resistant mechanism and improve soil environment play an important role.Isocitrate lyase structure prediction and gene sequence analysis, for the study of isocitrate lyase enzymology properties and metabolic mechanism of theory basis, for the Suillus luteus functions of mycorrhizal hypha growth and has important significance. The experiment using DNA sequencing technology and Rapid Amplifieation of cDNA ends technology obtained the isocitrate lyase cDNA and DNA sequence, amino acid sequence is analyzed, and forecasts the isocitrate lyase space structure.The results show that:According to the conserved sequence degenerate primers designed to Suillus luteus total RNA by PCR amplification of cDNA sequences 656bp in size. By using 3’RACE and 5’ RACE technique obtains the full-length cDNA sequences of 1852bp,this gene contains an open reading frame of 1620bp and encodes 539 amino acid protein sequence read.The homology of nucleotide sequence and Coniophora puteana, Laccaria hicolor, Coprinus cinereus, Schizophyllum and Agaricus bisporus were 76%,73%,72%,72%, 72%,the homology of amino acid sequence were 88%,88%,82%,81%,84%, And it analysis on ICL bioinformatics. PCR amplified DNA sequence, approximately 1928bp. Fragments of DNA sequence, it is found that it is consistent with the ICL gene encoding frame sequence, and the partial gene contains three introns, the length were 55 bp.49 bp,57bp and 49bp. Constructed pET28-ICL expression plasmid for further study isocitrate lyase enzymatic properties of the foundation.