Expression of long non-coding RNA MALAT1, NEAT1 and NEAT2 in peripheral blood of tuberculosis patients / 中华疾病控制杂志

Objective To analyze the differences in the expression levels of the lncRNA MALAT1, NEAT, NEAT2 in peripheral blood mononuclear cell (PBMC) from tuberculosis patients and healthy controls. Methods We detected the lncRNA expression levels in PBMC from 79 tuberculosis patients and 82 healthy controls by quantitative reverse transcription polymerase chain reaction, and analyzed the correlation between lncRNA expression levels and some clinical features and laboratory indicators in tuberculosis patients. Results The expression levels of MALAT1, NEAT1 in PBMC of tuberculosis patients were significantly higher than healthy controls (Z=-4.386, P<0.001; Z=-10.175, P<0.001). There was no significant difference in the expression of NEAT2 between tuberculosis patients and healthy controls (Z=-0.203，P=0.839). The correlation results of lncRNA levels and some clinical features, laboratory indicators in tuberculosis patients suggested that the NEAT2 level in PBMC of newly treated tuberculosis patients was higher than recurrent tuberculosis patients, while the NEAT2 level in PBMC of sputum smear positive tuberculosis patients was lower than that of sputum smear negative tuberculosis patients (all P<0.05). There was a negative correlation between MALAT1 level and erythrocyte sedimentation rate (rs=-0.256, P=0.034). Conclusion MALAT1 and NEAT1 are abnormally expressed in PBMC of tuberculosis patients, and may be involved in the pathogenesis of pulmonary tuberculosis.

Multipoint Mutation and Over-expression in Pichia pastoris of Phytase Gene / 中国生物工程杂志

According to bias in codon choice of Pichia pastoris, The phytase phyA gene from Aspergillus niger N25 was mutated without changing its amino acid sequence. The expression plasmid pPIC9k-phyAm was constructed and transformed into GS115 strain. Positive clones,of which the chromosomes were integrated with phyA gene,were identified by the phenotype and PCR. SDS-PAGE analysis suggust that the size of enzyme protein of the expression product was about 70.15kDa.Southern blotting analysis to the yeast transformants showed that phyA gene was intergrated into the chromosome genome. The phytase activity of PP-NP m-4-4 with codons optimized reached 136 000U/ml in malt wort culture medium after being induced with 36h, which was the 2.8 times of the original strain PP-NPm-8.