Tissue-Specific CRISPR/Cas9 System of Cotton Pollen with GhPLIMP2b and GhMYB24 Promoters

Lei Jianfeng,Dai Peihong,Li Jiyang,Yang Mi,Li Xiuqin,Zhang Wanqi,Zhou Guantong,WangzhenGuo,Liu Xiaodong 한국식물학회 2021 Journal of Plant Biology Vol.64 No.1

CRISPR/Cas9 technology is a powerful tool for improving crop genetic traits. However, CRISPR/Cas9 system for efficient expression of cotton germ cells has still not been established. In this study, we developed a tissue-specific vectors to drive Cas9 expression with GhPLIMP2b and GhMYB24 promoters and established the effective method to transform cotton pollen by Agrobacterium vacuum infiltration. GhPLIMP2b and GhMYB24 promoters of cotton pollen were cloned into Cas9 expression vectors. The sgRNAs targetting to CLA1, ERA1 and GGB (drought-resistant negative regulation genes) were designed and constructed into GhPLIMP2b and GhMYB24 promoter vectors. Cotton pollens were tranformed by Agrobacterium vacuum infiltration with GhPLIMP2b and GhMYB24 promoter vectors. The results of clone sequencing shown that mutation types of the sequence were mainly base substitutions wth the frequency from 3.29 to 6.45%. Eleven potential off-target sites were chosen and two sites were observed. Our results indicated that GhPLIM2bP::Cas9 and GhMYB24P::Cas9 editing vectors achieved targeted edition of endogenous genes in cotton pollen, but there were a few off-target effects. This study provides an effective gene-editing system for the rapid acquisition of cotton mutants using pollen as a transgenic receptor.

Cloning of porcine chemerin, ChemR23 and GPR1 and their involvement in regulation of Lipogenesis

( Jianfeng Huang ),( Jian Zhang ),( Ting Lei ),( Xiaodong Chen ),( Yan Zhang ),( Lulu Zhou ),( An Yu ),( Zhilong Chen ),( Ronghua Zhou ),( Zaiqing Yang ) 생화학분자생물학회 (구 한국생화학분자생물학회) 2010 BMB Reports Vol.43 No.7

Chemerin is a novel adipokine which is abundant in adipose tissue to promote adipocyte differentiation and with significant relativity to BMI and insulin sensitivity. We report here the molecular characterization of porcine chemerin and its receptors ChemR23 and GPR1, as well as their transcriptional regulation during lipogenesis. Chemerin was mainly expressed in liver, intestine, kidney and adipose tissue, consistent with the expression pattern of GPR1, but not ChemR23, which was predominantly present in spleen and temperately in adipose tissue. We further investigated the lipogenesis-related transcriptional activation of PPARγ and KLF15 on chemerin and its receptors. The data showed that KLF15, but not PPARγ, can up-regulate the mRNA level of chemerin, ChemR23 and GPR1, which was consistent with the results of luciferase assay that confirmed the effect of KLF15 on ChemR23 promoter. Taken together, our data provide basic molecular information for the further investigation on the function of chemerin in lipogenesis. [BMB reports 2010; 43(7): 491-498]

Robust Estimation of Angular Parameters of the Surface Moving Basin Boundary Induced by Coal Mining: A Case of Huainan Mining Area

Lei Wang,Chuang Jiang,Tao Wei,Nan Li,Shenshen Chi,Jianfeng Zha,Suyang Fang 대한토목학회 2020 KSCE JOURNAL OF CIVIL ENGINEERING Vol.24 No.1

The data of surface movement observation points could be used to obtain the synthetic angles of draw and critical deformation of surface movement basin in the mining area with unconsolidated thick layers. However, they couldn’t be used to get the angles of draw and critical deformation of unconsolidated layers and rock layers, which are called angular parameters of surface movement basin and can be used to describe the outermost boundary and the dangerous boundary. Therefore, the paper presented an estimation method of angular parameters of surface moving basin based on least squares of Huber selecting weight iteration (LSHSWI). The method firstly classified the surface movement observation points with similar geological and mining conditions as the same kinds. Then, the synthetic angles of critical deformation and draw obtained by the observation points were taken as observation values, and angular parameters of surface movement basin were taken as unknown parameters.According to the geometric relationship between the synthetic angles and the angular parameters, the observation equation was established. Finally, the parameters estimation method based on LSHSWI was constructed with consideration of errors in the synthetic angles of draw and critical deformation. The simulation experiments show that the LSHSWI estimation method of angular parameters of surface movement basin has a strong anti-interference ability to errors of observation values. The engineering application shows that the parameter estimation method based on LSHSWI is scientific and practical.

The effect of Glomus intraradices on the physiological properties of Panax ginseng and on rhizospheric microbial diversity

Lei Tian,Shaohua Shi,Lina Ma,Xue Zhou,Shasha Luo,Jianfeng Zhang,Baohui Lu,Chunjie Tian 고려인삼학회 2019 Journal of Ginseng Research Vol.43 No.1

Background: Glomus intraradices is a species of arbuscular mycorrhizal fungi that, as an obligate endomycorrhiza, can form mutually beneficial associations with plants. Panax ginseng is a popular traditional Chinese medicine; however, problems associated with ginseng planting, such as pesticide residues, reduce the ginseng quality. Methods: In this experiment, we studied the effect of inoculating G. intraradices on several physiological properties and microbial communities of ginseng. UV-Visible Spectrum method was used to detect physical properties. Denaturing gradient gel electrophoresis method was used to analyze microbial communities. Results: The results indicated that inoculation with G. intraradices can improve the colonization rate of lateral ginseng roots, increase the levels of monomeric and total ginsenosides, and improve root activity as well as polyphenol oxidase and catalase activities. We also studied the bacterial and fungal communities in ginseng rhizospheric soil. In our study, G. intraradices inoculation improved the abundance and Shannon diversity of bacteria, whereas fungi showed a reciprocal effect. Furthermore, we found that G. intraradices inoculation might increase some beneficial bacterial species and decreased pathogenic fungi in rhizospheric soil of ginseng. Conclusion: Our results showed that G. intraradices can benefit ginseng planting which may have some instructive and practical significance for planting ginseng in farmland.

The effect of Glomus intraradices on the physiological properties of Panax ginseng and on rhizospheric microbial diversity

Tian, Lei,Shi, Shaohua,Ma, Lina,Zhou, Xue,Luo, Shasha,Zhang, Jianfeng,Lu, Baohui,Tian, Chunjie The Korean Society of Ginseng 2019 Journal of Ginseng Research Vol.43 No.1

Background: Glomus intraradices is a species of arbuscular mycorrhizal fungi that, as an obligate endomycorrhiza, can form mutually beneficial associations with plants. Panax ginseng is a popular traditional Chinese medicine; however, problems associated with ginseng planting, such as pesticide residues, reduce the ginseng quality. Methods: In this experiment, we studied the effect of inoculating G. intraradices on several physiological properties and microbial communities of ginseng. UV-Visible Spectrum method was used to detect physical properties. Denaturing gradient gel electrophoresis method was used to analyze microbial communities. Results: The results indicated that inoculation with G. intraradices can improve the colonization rate of lateral ginseng roots, increase the levels of monomeric and total ginsenosides, and improve root activity as well as polyphenol oxidase and catalase activities. We also studied the bacterial and fungal communities in ginseng rhizospheric soil. In our study, G. intraradices inoculation improved the abundance and Shannon diversity of bacteria, whereas fungi showed a reciprocal effect. Furthermore, we found that G. intraradices inoculation might increase some beneficial bacterial species and decreased pathogenic fungi in rhizospheric soil of ginseng. Conclusion: Our results showed that G. intraradices can benefit ginseng planting which may have some instructive and practical significance for planting ginseng in farmland.

Aα-SPECTRAL EXTREMA OF GRAPHS WITH GIVEN SIZE AND MATCHING NUMBER

Xingyu Lei,Shuchao Li,Jianfeng Wang 대한수학회 2023 대한수학회보 Vol.60 No.4

In 2017, Nikiforov proposed the $A_{\alpha}$-matrix of a graph $G$. This novel matrix is defined as $$A_{\alpha}(G)=\alpha D(G)+(1- \alpha )A(G),~\alpha \in [0,1],$$ where $D(G)$ and $A(G)$ are the degree diagonal matrix and adjacency matrix of $G$, respectively. Recently, Zhai, Xue and Liu \cite{Zhai2020} considered the Brualdi-Hoffman-type problem for $Q$-spectra of graphs with given matching number. As a continuance of it, in this contribution we consider the Brualdi-Hoffman-type problem for $A_{\alpha}$-spectra of graphs with given matching number. We identify the graphs with given size and matching number having the largest $A_{\alpha}$-spectral radius for $\alpha \in [\frac{1}{2},1)$.

Efficient Expression and Purification of Recombinant Alcohol Oxidase in Pichia pastoris

Yunping Liu,Jianfeng Pan,Peilian Wei,Jianzhong Zhu,Lei Huang,Jin Cai,Zhinan Xu 한국생물공학회 2012 Biotechnology and Bioprocess Engineering Vol.17 No.4

In order to improve the production of alcohol oxidase (AOX), a recombinant Pichia pastoris (P. pastoris)system was constructed by transformation of the plasmid pPIC9K-AOX into P. pastoris GS115. The effects of different expression conditions on alcohol oxidase activity in the culture supernatant were investigated in the shake flask scale. The results showed that the highest extracellular activity (562 U/L) of alcohol oxidase was obtained after 56h induction with 4% methanol at OD600 1.0 in the medium containing 50 g/L maltose, which is about 4.2 folds higher than previously reported. High-purity functional recombinant AOX (>90%) was purified from the culture with the Ni-NTA affinity column and Sephadex G-100 chromatographical methods, with a total recovery rate of 68.9%. Further studies showed that the purified rAOX had similar enzymatic characteristics as the native enzyme, except that the thermal stability and resistance to H2O2 inhibition of rAOX were significantly greater compared to the previous report. The purified rAOX was well tolerant to various water-miscible organic solvents. This efficient expression and purification process will be promising for large-scale production of rAOX as an important diagnostic enzyme for alcohol detection in many areas.

Restoration of dysregulated intestinal barrier and inflammatory regulation through synergistically ameliorating hypoxia and scavenging reactive oxygen species using ceria nanozymes in ulcerative colitis

Ying Zhang,Hengyu Lei,Pengchong Wang,Qinyuan Zhou,Jie Yu,Xue Leng,Ruirui Ma,Danyang Wang,Kai Dong,Jianfeng Xing,Yalin Dong 한국생체재료학회 2023 생체재료학회지 Vol.27 No.00

Background Reactive oxygen species (ROS) overproduction and excessive hypoxia play pivotal roles in the initiation and progression of ulcerative colitis (UC). Synergistic ROS scavenging and generating O2 could be a promising strategy for UC treatment. Methods Ceria nanozymes (PEG-CNPs) are fabricated using a modified reverse micelle method. We investigate hypoxia attenuating and ROS scavenging of PEG-CNPs in intestinal epithelial cells and RAW 264.7 macrophages and their effects on pro-inflammatory macrophages activation. Subsequently, we investigate the biodistribution, pharmacokinetic properties and long-term toxicity of PEG-CNPs in mice. PEG-CNPs are administered intravenously to mice with 2,4,6-trinitrobenzenesulfonic acid-induced colitis to test their colonic tissue targeting and assess their anti-inflammatory activity and mucosal healing properties in UC. Results PEG-CNPs exhibit multi-enzymatic activity that can scavenge ROS and generate O2, promote intestinal epithelial cell healing and inhibit pro-inflammatory macrophage activation, and have good biocompatibility. After intravenous administration of PEG-CNPs to colitis mice, they can enrich at the site of colonic inflammation, and reduce hypoxia-induced factor-1α expression in intestinal epithelial cells by scavenging ROS to generate O2, thus further promoting disrupted intestinal mucosal barrier restoration. Meanwhile, PEG-CNPs can effectively scavenge ROS in impaired colon tissues and relieve colonic macrophage hypoxia to suppress the pro-inflammatory macrophages activation, thereby preventing UC occurrence and development. Conclusion This study has provided a paradigm to utilize metallic nanozymes, and suggests that further materials engineering investigations could yield a facile method based on the pathological characteristics of UC for clinically managing UC.

ZNRF1 can inhibit proliferation and stemness properties of leukemia NB4 cells

Yuechao Zhao,TongJuan Li,Lei Zhao,Jue Wang,Zhen Shang,Wei Huang,Jianfeng Zhou 한국통합생물학회 2016 Animal cells and systems Vol.20 No.4

As a newly discovered E3 ubiquitin ligase, ZNRF1 protein participates in controlling Wallerian degeneration, but the role in cancer biology has been seldom reported. We analyzed the change of proliferation, apoptosis, stemness and protein levels of AKT and STAT5 after leukemia NB4 cells were treated with ZNRF1 siRNA and with expression vector. The results showed that overexpression of ZNRF1 inhibited proliferation and increased apoptosis of NB4 cells, but ZNRF1 siRNA did not influence the proliferation and apoptosis. Over-expression of ZNRF1 diminished cancer stem cell properties and down-regulated AKT and STAT5. This is the first study to show that ZNRF1 inhibited proliferation and stemness properties of leukemia NB4 cells possibly through AKT signal and STAT5 signal.

A Centralized Displacement Operation with Application to Artifact Reduction in Ultrasonic Elastography

Dangguo Shao,Yong Chen,Sanli Yi,Lei Ma,Jianfeng He,Dong C. Liu 보안공학연구지원센터(IJSIP) 2014 International Journal of Signal Processing, Image Vol.7 No.2