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Establishment of hybridization breeding of Ganoderma lucidum by protoplast monokaryogenesis method
Wu Jiaojiao,Fu Lizhong,Wu Xueqian,Xu Xiuhong,Li Haibo,Wu Qingqi,Wei Hailong,Cheng Junwen 한국버섯학회 2010 한국버섯학회지 Vol.8 No.4
Four Ganoderma lucidum strains, Chizhi 05, Jingda, Huizhou and Xinzhou, were screened out as hybrid parent in order to establish G. lucidum cross breeding system that based on protoplast monokaryogenesis method. Monokaryotic strains of each parental strains were obtained and mating type of each monokaryotic strains were determined. One to three monokaryotic strains that have different mating types were mated, and hybrids were identified by clamp connection observation and antagonist response. The results showed that the number of monokaryon came from Chizhi 05, Jingda, Huizhou and Xinzhou was 9, 14, 40 and 38, respectively. Only one mating type was obtained from Jingda, and two mating types were obtained from the other three strains, Chizhi 05, Huizhou and Xinzhou, respectively. Chi-square test showed that the ratio of two mating types of the three strains was 1:1. Fourteen monokaryotic strains of different mating types from 4 parental strains were select as a cross- breeding materia, and 17 hybrids were obtained, which were identified by clamp connection observation and antagonist response. This study proclaimed that the practicality of the hybridization breeding of G. lucidum by protoplast monokaryogenesis method.
Han Wu,Xiuhong Wang,Tingting Wu,Su Yang 한국유전학회 2020 Genes & Genomics Vol.42 No.3
Background Dysregulation of miR-489 in human tumors has been widely reported. Lactate dehydrogenase isoform A (LDHA)-mediated aerobic glycolysis participates in proliferation of multiple myeloma (MM) cells. Objective To investigate whether miR-489 induced MM growth inhibition via targeting to LDHA-mediated aerobic glycolysis. Methods Expression of miR-489 in representative MM cell lines was determined via qRT-PCR (quantitative real-time polymerase chain reaction). MTT (3-(4, 5-di methyl thiazol-2-yl)-2, 5-di phenyl tetrazolium bromide) and colony formation assays were utilized to detect cell viability and proliferation. Effect of miR-489 on aerobic glycolysis was detected via glucose uptake, lactate and ATP production. Binding ability between miR-489 and LDHA was conducted via luciferase activity assay. Results MiR-489 was down-regulated in representative MM cell lines. Gain-of functional assays indicated that over-expression of miR-489 decreased cell viability and inhibited cell proliferation of MM cells. Moreover, miR-489 inhibited aerobic glycolysis via decrease of glucose uptake, lactate and ATP production. LDHA was identified as target of miR-489, suggesting a negative correlation between miR-489 and LDHA in MM cells. Mechanically, the inhibition ability of miR-489 on proliferation of MM cells was through inhibition of LDHA-mediated aerobic glycolysis. Conclusions miR-489 inhibited MM tumor growth via LDHA-mediated glycolytic metabolism, suggesting potential therapeutic target ability of miR-489/LDHA for MM.
( Jian Tu ),( Kezong Qi ),( Ting Xue ),( Haiting Wei ),( Yongzheng Zhang ),( Yanli Wu ),( Xiuhong Zhou ),( Xiaolong Lv ) 한국미생물 · 생명공학회 2015 Journal of microbiology and biotechnology Vol.25 No.12
Avian beta-defensin 9 (AvBD9) is a small cationic peptide consisting of 41 amino acids that plays a crucial rule in innate immunity and acquired immunity in chickens. Owing to its wide antibacterial spectrum, lack of a residue, and failure to induce bacterial drug resistance, AvBD9 is expected to become a substitute for conventional antibiotics in the livestock and poultry industries. Using the preferred codon of Pichia pastoris, the mature AvBD9 peptide was designed and synthesized, based on the sequence from GenBank. The P. pastoris constitutive expression vector pGHKα was used to construct a pGHKα-AvBD9 recombinant plasmid. Restriction enzyme digestion was performed using SacI and BglII to remove the ampicillin resistance gene, and the plasmid was electrotransformed into P. pastoris GS115. High-expression strains with G418 resistance were screened, and the culture supernatant was analyzed by Tricine-SDS-PAGE and western blot assay to identify target bands of about 6 kDa. A concentrate of the supernatant containing AvBD9 was used for determination of antimicrobial activity. The supernatant concentrate was effective against Escherichia coli, Salmonella paratyphi, Salmonella pullorum, Pseudomonas aeruginosa, Enterococcus faecalis, and Enterobacter cloacae. The fermentation product of P. pastoris carrying the recombinant AvBD9 plasmid was adjusted to 1.0 × 108 CFU/ml and added to the drinking water of white feather broilers at different concentrations. The daily average weight gain and immune organ indices in broilers older than 7 days were significantly improved by the AvBD9 treatment.
( Yanling Quan ),( Lin Wang ),( Yisheng Liu ),( Jingxiang Cong ),( Shengquan Xie ),( Xiuhong Wu ) 한국화학공학회 2015 Korean Chemical Engineering Research(HWAHAK KONGHA Vol.53 No.3
Plackett-Burman (PB) and Box-Behnken (BB) experimental designs were used to optimize fermentation variables for the biotransformation of glycyrrhizin (GL) to monoglucuronyl-glycyrrhetinic acid (MGGA). The PB design was first used to screen the important factors among the medium variables. The steepest ascent method was used to approach the optimum range for each of these factors. The BB design was finally used to analyze the response surfaces of the screened factors for further optimization. The optimized conditions for this system were 0.7 g/L MgSO4·7H2O, 1.19 g/L GL, and cultivation for six days. The biotransformation of GL to MGGA could reach up to 35.72%, which is a good result for this kind of transformation.