The antifungal protein HAS1 is a new antifungal protein isolated from the genome of Bacillus subtilis HAS,which has a good inhibitory effect on various pathogenic fungi in sugarcane.This study aimed at evaluating the ...The antifungal protein HAS1 is a new antifungal protein isolated from the genome of Bacillus subtilis HAS,which has a good inhibitory effect on various pathogenic fungi in sugarcane.This study aimed at evaluating the immunological transfer reaction of the test sample through repeated skin contact by observing whether the cavy skin repeatedly exposed to purified protein HAS1 suffers from allergic reaction and how strong is the allergic reaction.The results showed that the test group and the vehicle control group exhibited no allergic reaction in the skin immediately and at 24,48 and 72 h and had an mean reaction score and a sensitization rate both of 0,so the results of the naked eye observation were both nonallergenic to cavy skin;and in the positive control group,the cavies were observed to be highly sensitized immediately after removing the drug,mildly sensitized at 24 h,and not sensitized at 48 and 72 h,which meant the skin allergy was alleviated with the observation time.It is suggested that the purified protein HAS1 is negative for cavy skin anaphylaxis test (nonallergenic to cavy skin),which provides an experimental basis for further utilization of the protein and its coding gene.展开更多
The bglS gene encoding endo-1,3-1,4-β-glucanase from Bacillus subtil＆ was cloned and sequenced in this study. The bglS expression cassette, including PGK1 promoter, bglS gene fused to the signal sequence of the yeas...The bglS gene encoding endo-1,3-1,4-β-glucanase from Bacillus subtil＆ was cloned and sequenced in this study. The bglS expression cassette, including PGK1 promoter, bglS gene fused to the signal sequence of the yeast mating pheromone a-factor （MFals）, and ADH1 terminator with G418-resistance as the selected marker, was constructed. Then one of the PEP4 allele of Saccharomyces cerevisiae WZ65 strain was replaced by bglS expression cassette using chromosomal integration of polymerase chain reaction （PCR）-mediated homologous recombination, and the bglS gene was expressed simultaneously. The recombinant strain S. cerevisiae （SC-βG） was preliminarily screened by the clearing hydrolysis zone formed after the barley β-glucan was hydrolyzed in the plate and no proteinase A （PrA） activity was measured in fermenting liquor. The results of PCR analysis of genome DNA showed that one of the PEP4 allele had been replaced and bglS gene had been inserted into the locus of PEP4 gene in recombinant strains. Different endo-1,3-1,4-β-glucanase assay methods showed that the recombinant strain SC-βG had high endo-1,3-1,4-β-glucanase expression level with the maximum of 69.3 U/（h·ml） after 60 h of incubation. Meanwhile, the Congo Red method was suitable for the determination of endo-1,3-1,4-β-glucanase activity during the actual brewing process. The current research implies that the constructed yeast strain could be utilized to improve the industrial brewing property of beer.展开更多
Limb and CNS expressed 1 like(LIX1L) is over-expressed in several types of tumors.However,the function of LIX1L in glucose metabolism and hepatocellular carcinoma(HCC) progression remains elusive.Here we report that L...Limb and CNS expressed 1 like(LIX1L) is over-expressed in several types of tumors.However,the function of LIX1L in glucose metabolism and hepatocellular carcinoma(HCC) progression remains elusive.Here we report that LIX1L is over-expressed in human HCC tissues,which predicts unfavorable prognosis.LIX1L deficiency in vivo significantly attenuated liver cancer initiation in mice.Functional studies indicated that LIX1L overexpression elevated proliferation,migratory,invasive capacities of HCC cells in vitro,and promoted liver cancer growth and metastasis in vivo.LIX1L knockdown up-regulated fructose-1,6-bisphosphatase(FBP1) expression to reduce glucose consumption as well as lactate production.Mechanistically,LIX1L increased miR-21-3p expression,which targeted and suppressed FBP1,thereby promoting HCC growth and metastasis.MiR-21-3p inhibitor could abrogate LIX1L induced enhancement of cell migration,invasion,and glucose metabolism.Inhibition of miR-21-3p suppressed tumor growth in an orthotopic tumor model.Our results establish LIX1L as a critical driver of hepatocarcinogenesis and HCC progression,with implications for prognosis and treatment.展开更多
基金Supported by National Natural Science Foundation(31471555)National Key Research and Development Project(SQ2018YFD020024)
文摘The antifungal protein HAS1 is a new antifungal protein isolated from the genome of Bacillus subtilis HAS,which has a good inhibitory effect on various pathogenic fungi in sugarcane.This study aimed at evaluating the immunological transfer reaction of the test sample through repeated skin contact by observing whether the cavy skin repeatedly exposed to purified protein HAS1 suffers from allergic reaction and how strong is the allergic reaction.The results showed that the test group and the vehicle control group exhibited no allergic reaction in the skin immediately and at 24,48 and 72 h and had an mean reaction score and a sensitization rate both of 0,so the results of the naked eye observation were both nonallergenic to cavy skin;and in the positive control group,the cavies were observed to be highly sensitized immediately after removing the drug,mildly sensitized at 24 h,and not sensitized at 48 and 72 h,which meant the skin allergy was alleviated with the observation time.It is suggested that the purified protein HAS1 is negative for cavy skin anaphylaxis test (nonallergenic to cavy skin),which provides an experimental basis for further utilization of the protein and its coding gene.
基金the National Hi-Tech Research and Develop-ment Program (863) of China (No. 2007AA10Z315)the Natural Science Foundation of Zhejiang Province, China (No. Z304076)
文摘The bglS gene encoding endo-1,3-1,4-β-glucanase from Bacillus subtil＆ was cloned and sequenced in this study. The bglS expression cassette, including PGK1 promoter, bglS gene fused to the signal sequence of the yeast mating pheromone a-factor （MFals）, and ADH1 terminator with G418-resistance as the selected marker, was constructed. Then one of the PEP4 allele of Saccharomyces cerevisiae WZ65 strain was replaced by bglS expression cassette using chromosomal integration of polymerase chain reaction （PCR）-mediated homologous recombination, and the bglS gene was expressed simultaneously. The recombinant strain S. cerevisiae （SC-βG） was preliminarily screened by the clearing hydrolysis zone formed after the barley β-glucan was hydrolyzed in the plate and no proteinase A （PrA） activity was measured in fermenting liquor. The results of PCR analysis of genome DNA showed that one of the PEP4 allele had been replaced and bglS gene had been inserted into the locus of PEP4 gene in recombinant strains. Different endo-1,3-1,4-β-glucanase assay methods showed that the recombinant strain SC-βG had high endo-1,3-1,4-β-glucanase expression level with the maximum of 69.3 U/（h·ml） after 60 h of incubation. Meanwhile, the Congo Red method was suitable for the determination of endo-1,3-1,4-β-glucanase activity during the actual brewing process. The current research implies that the constructed yeast strain could be utilized to improve the industrial brewing property of beer.
基金supported by National Natural Science Foundation of China (No. 82074068 and 81872889)Natural Science Foundation of Jiangsu Province (BK20181332, China) to Hao Zhang+1 种基金The Drug Innovation Major Project (2018ZX09711-001007 and 2018ZX09735002-003, China)the “Double First-Class” University Project (CPU2018GF03, China) to Lingyi Kong。
文摘Limb and CNS expressed 1 like(LIX1L) is over-expressed in several types of tumors.However,the function of LIX1L in glucose metabolism and hepatocellular carcinoma(HCC) progression remains elusive.Here we report that LIX1L is over-expressed in human HCC tissues,which predicts unfavorable prognosis.LIX1L deficiency in vivo significantly attenuated liver cancer initiation in mice.Functional studies indicated that LIX1L overexpression elevated proliferation,migratory,invasive capacities of HCC cells in vitro,and promoted liver cancer growth and metastasis in vivo.LIX1L knockdown up-regulated fructose-1,6-bisphosphatase(FBP1) expression to reduce glucose consumption as well as lactate production.Mechanistically,LIX1L increased miR-21-3p expression,which targeted and suppressed FBP1,thereby promoting HCC growth and metastasis.MiR-21-3p inhibitor could abrogate LIX1L induced enhancement of cell migration,invasion,and glucose metabolism.Inhibition of miR-21-3p suppressed tumor growth in an orthotopic tumor model.Our results establish LIX1L as a critical driver of hepatocarcinogenesis and HCC progression,with implications for prognosis and treatment.
