Mycobacterium tuberculosis h37rv encoding gene and its application
A technology of Mycobacterium tuberculosis, encoding gene, applied in application, genetic engineering, plant genetic improvement and other directions, can solve the problems of wrong gene boundary, complicated operation, long cycle, etc., to achieve the effect of easy comparison
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Embodiment 1
[0033]Example 1: Searching for missing annotation coding genes in the H37Rv strain genome
[0034] 1.1 High-coverage proteome verification of the H37Rv strain genome
[0035] A deep coverage study of the proteome was performed on the H37Rv strain using high-coverage proteome technology. Based on the Tuberculosis (20160307) database, the genome was annotated and encoded using the pFind 3 engine. In order to discover new protein coding regions, based on proteomics technology, we used pAnno software to translate the six-reading frame database of H37Rv's genome (NC_000962.3) file published by NCBI, and used this database to perform mass spectrometry data for new peptides. Segments and identification of new proteins. In order to reduce the false positive rate, we used three filtering methods for separately estimating category FDR for annotated peptides and new peptides during the data filtering process, namely S-FDR, T-FDR I and T-FDRII.
[0036] After data analysis, we identifi...
Embodiment 2
[0051] Embodiment 2: Establish the method for identifying MTBC complex group
[0052] (1) Design primers:
[0053] Based on the CDS sequence of the Rv3202B (+|3580526-3580684|) gene shown in SEQ ID NO.1, PCR primers were designed using Oligo7.0. The primer sequences are as follows:
[0054] F: 5'-TCATCGACCTTCACGTACA-3' (SEQ ID NO.4);
[0055] R: 5'-AAAGTGATGGGCTAACCG-3' (SEQ ID NO.5)
[0056] The positional relationship between the above primers and the Rv3202B (+|3580526-3580684|) gene is shown below, where the corresponding positions of the primers are underlined.
[0057] GTGA TCATCGACCTTCACGTACA GCGCTACGGCCCGTCAGGGCCCGCGCGGGTGCTGACCATCCACGGAGTGACCGAGCACGGGCGCATCTGGCA CCGGTTAGCCCATCACTTT GCCCGAAATCCCCATCGCCGCACCCGATCTGCTGGGCCACGGTAG (SEQ ID NO. 1)
[0058] (2) Extract the total DNA of the strains to be tested including M.tuberculosis H37Rv, 40 standard strains of Mycobacterium are preserved by the China Medical Bacteria Culture Collection Center (CMCC), and the remai...
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