Title: Cloning and analyses of the protease gene from Bacillus subtilis strain ZJ06 for milk-protein degradation
Abstract:Cloning of protease gene for milk-protein degradation from Bacillus subtilis strain ZJ06,analyzing the structure and predicting the characters of protease gene.A fragment encoding an alkaline protease...Cloning of protease gene for milk-protein degradation from Bacillus subtilis strain ZJ06,analyzing the structure and predicting the characters of protease gene.A fragment encoding an alkaline protease was amplified by PCR from genomic DNA of Bacillus subtilis strain ZJ06,and then cloned cloning plasmid for sequencing analysis.The specific products were amplified using cDNA derived from total RNA of Bacillus subtilis strain ZJ06 in RT-PCR.In addition,colony in situ hybridization with random primed DNA labeling with digoxigenin-dUTP also indicated this gene was confirmed.A new gene encoding an alkaline protease(apr) was cloned and its nucleotide sequence was determined(GenBank accession number EU386607).DNA sequence analysis revealed a single open reading frame of 1146 bp encoding 382 amino acids with the calculated molecular mass of 39 ku and the signal peptide was 30 amino acids.Amino acid analysis indicated this gene is belonging to secreted protease with zymogen of 352 amino acids and mature protease of 275 amino acids.The percentage of GC in the mature protease was 51.4% and its calculated molecular mass was 27.4 ku.The expression of the alkaline protease gene was confirmed by RT-PCR and colony in situ hybridization.Read More
Publication Year: 2010
Publication Date: 2010-01-01
Language: en
Type: article
Access and Citation
AI Researcher Chatbot
Get quick answers to your questions about the article from our AI researcher chatbot