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Table of Content

    • Current Biotechnology
      Volume 11 Issue 1 25 January 2021
    • Application Progress of New Gene Editing Technology in Single Cell Microalgae
    • CAO Haohao1, ZHANG Hongbing1*, XUE Xifa1, LI Zuoqun2, YAN Hongbo1, LI Huixuan1
    • 2021, 11(1):  9-15.  DOI: 10.19586/j.2095-2341.2020.0090
    • Abstract ( ) PDF (3848KB)( )
      • Microalgae, as single-celled organisms, are widely distributed in oceans and freshwater lakes. Microalgae are  used in the production of bioenergy and high-value products because they can accumulate a large amount of biomass in cells during their growth. Although microalgae have great potential, bottlenecks such as low lipid rate have not been solved in existing research. In recent years, gene engineering technology, especially new gene editing technology, has developed rapidly, and genome editing technology is fast and effective, which is conducive to fully understanding the genetic and molecular biological information of microalgae. Therefore, its application in microalgae has become a hot field in microalgae genetic engineering and bioengineering, and has attracted extensive attention. Based on this, several new gene editing technologies such as zinc finger nuclease (ZFN), transcription activator-like effector nuclease (TALEN) and clustered regularly spaced short palindromic repeats (CRISPR)/CAS (CRISPR-associated systems) nuclease system, etc, were briefly introduced. The application progress of these technologies in microalgae was reviewed, and the prospect of microalgae gene editing was made, in order to provide reference for further understanding and solving the problems related to microalgae gene editing.
    • #br# Cyclic RNA and its Application in Ecotoxicology
    • WANG Rui1, LI Liang2
    • 2021, 11(1):  16-25.  DOI: 10.19586/j.2095-2341.2020.0095
    • Abstract ( ) PDF (2114KB)( )
      • Circular RNAs (circRNAs) are a class of covalent closed ring noncoding RNA, which has the characteristics of conservative evolution, stable structure and tissue specific expression. CircRNAs can act as miRNAs sponges to affect the regulation of genes, and interact with RNA binding proteins (RBPs). Some studies have shown that some circRNAs have the potential to be translated into proteins. CircRNAs has been proved to have specificity, stability and regulatory function for some diseases, such as cancer, diabetes, cardiovascular diseases, neurodegenerative diseases, etc. They can be used as potential biomarkers for diagnosis and prognosis and therapeutic targets. Recently, it has been found that circRNAs are involved in the occurrence and development of toxic effects induced by environmental chemical pollutants. At present, the toxicity endpoint of evaluating the relationship between chemical pollutants and toxic effects of environmental pollutants in ecotoxicology research is usually affected by genetic polymorphism and epigenetics. Considering the phenomenon of differential expression of circRNAs in organisms after exposure to environmental chemical pollutants, circRNAs may also be used as biomarkers in ecotoxicological researches. Based on the above, the biosynthesis and degradation, biological functions, analytical methods of circRNAs and their current applications in ecotoxicology were reviewed, and the application of circRNAs as molecular biomarkers in early diagnosis and ecological risk assessment of environmental pollutant exposure was prospected, in order to provide reference for ecotoxicological researches and environmental risk assessment.
    • Progress on Structure and Function of FAM96A and FAM96B
    • ZHANG Didi, GU Yuxuan, SUN Xiaolin, ZHANG Lina*
    • 2021, 11(1):  26-32.  DOI: 10.19586/j.2095-2341.2020.0092
    • Abstract ( ) PDF (2097KB)( )
      • Family with sequence similarity 96 member A and B (FAM96A and FAM96B) are two highly conserved homologous proteins belonging to MIP18 (MMS 19-interacting protein of 18 kD) family. MIP18 is a subunit of MMDX(MMS19-MIP18-XPD) complex related to mitotic spindle. Studies have shown that the expressions of FAM96A and FAM96B in human gastrointestinal stromal tumors, colon cancer, liver cancer, gastric cancer and breast cancer are significantly decreased compared with corresponding normal tissues, suggesting that they may participate in the occurrence and development of tumors as potential tumor suppressor genes. However, the molecular mechanisms of FAM96A and FAM96B in the occurrence and development of tumors are not very clear at present. In addition, it was found that FAM96A and FAM96B could play a variety of different functions in vivo by interacting with other different proteins. Therefore, the research progress on the structure and function of FAM96A and FAM96B was reviewed and summarized, and the research prospect of molecular mechanism and identification of interacting proteins in tumor development was prospected, in order to lay a foundation for clinical use of FAM96A and FAM96B as new tumor diagnostic markers and therapeutic targets, and provide a basis for revealing more new functions of FAM96A and FAM96B in vivo.
    • Research Progress of JAK2/STAT3 Served as a Novel Anticancer Drug Target
    • ZHANG Wenying1,2§, WANG Siqing1§, ZHANG Xinyan, LI Qingwei1,2,3, LI Yingying1,2,3*
    • 2021, 11(1):  33-39.  DOI: 10.19586/j.2095-2341.2020.0059
    • Abstract ( ) PDF (2137KB)( )
      • Compared with conventional chemotherapy, targeted drug therapy can better distinguish normal cells from tumor cells, and then kill tumor cells more specifically without damaging normal cells. As a signal transduction pathway stimulated by cytokines, Janus kinases/signal transducer and activator of transcription (JAK/STAT) signaling pathway is involved in many important biological processes, such as cell proliferation, differentiation and immune regulation. JAK2/STAT3 signaling pathway is an important component of JAK/STAT signaling pathway, which has been proved to be over-activated in a variety of tumor cells, particularly in some solid tumor cells, so it is expected to become a new target of anticancer drugs. Based on this, the roles of JAK2/STAT3 signaling pathway molecules in liver cancer, breast cancer and pancreatic cancer were reviewed, and then expounded from the perspectives of JAK2/STAT3 signaling pathway molecules participating in tumor cell proliferation, inducing apoptosis and mediating invasion and metastasis, so as to provide theoretical guidance for improving the effect of cancer precision therapy and open up new approaches for cancer treatment.
    • Research Progress on the Endoplasmic Reticulum Stress and its Mechanism in Diabetic Nephropathy
    • JIN Tong, CHEN Cheng
    • 2021, 11(1):  40-46.  DOI: 10.19586/j.2095-2341.2020.0086
    • Abstract ( ) PDF (2680KB)( )
      • Diabetic nephropathy (DN) is the most common microvascular complication of diabetes and one of the main causes of secondary renal disease leading to end-stage renal disease (ESRD). A variety of factors such as lack of oxygen, oxidative stress, virus infection, genetic mutations can lead to an imbalance in the homeostasis of the endoplasmic reticulum, and the accumulation of a large number of unfolded and misfolded proteins, namely the formation of endoplasmic reticulum stress (ERS), thereby activating the unfolded protein response (UPR) mediated three classic adaptive response pathways to restore endoplasmic reticulum homeostasis and cell viability. However, if the stimulation is too strong or persistent, the apoptotic signaling pathway will be activated. A large number of studies have shown that ERS is related to the occurrence and development of DN and is involved in the process of different types of renal cell injury. Therefore, ERS as an effective target for the treatment of DN has a very important research prospect. Therefore, the regulation of  ERS may provide a new theoretical support for the treatment of DN. In this paper, ERS associated signaling pathways and their roles in DN were reviewed, which was expected to provide reference for DN treatment research.
    • Functional Study on Ferredoxin-like Protein Slr1205 in Synechocystis sp. PCC 6803
    • DONG Jie, ZHANG Hao, ZHANG Pengpeng*
    • 2021, 11(1):  69-78.  DOI: 10.19586/j.2095-2341.2020.0081
    • Abstract ( ) PDF (7064KB)( )
      • Ferredoxin-like (Fd-like) proteins are involved in many important physiological roles in plants, such as regulation of chloroplast development. However, their effects in cyanobacteria have not revealed. In this work, the function of Synechocystis Fd-like protein Slr1205 was analyzed by comparison of physiological and biochemical properties of Δslr1205 and WT grown under different carbon sources and photoperiod. It showed that the growth rate of Δslr1205 was lower than that of the WT under conditions of high CO2 concentration autotrophy, mixotrophy and photoheterotrophy, while no significant difference was observed under air-level CO2 autotrophic cultures. In agreement with these, Δslr1205 cells decreased the respiratory rate and accumulation of NDH-1L complex, an enzyme closely related to cellular respiration in cyanobacteria. In all our tested conditions, Δslr1205 had higher carotenoid content and yellowish phenotype compared to WT. These data suggested that lack of Fd-like protein Slr1205 led to low growth rate at adequate carbon supply, which was probably due to insufficient energy caused by down-regulation of respiration. The work provided not only the basis for further investigation of cyanobacterial Fd-like proteins, but also a new direction for understanding of regulation mechanisms of photosynthesis and respiration.
    • Analysis of Microbial Diversity of Skin Mucus and Intestinal Contents in Hexagrammos otakii Based on 16S rRNA High-throughput Sequencing Technology
    • FAN Ying, YU Xiaoqing, LI Le, WANG Xiaolu, YE Haibin, HU Fawen, DIAO Jing*, LIU Hongjun
    • 2021, 11(1):  79-90.  DOI: 10.19586/j.2095-2341.2020.0078
    • Abstract ( ) PDF (4633KB)( )
      • Hexagrammos otakii is a kind of representative cold water fish with high value. Based on the analysis about microflora of the skin mucus and intestinal contents in H. otakii, the diversity of the microflora and the potential pathogenic microorganisms were studied. The genomic DNA of skin mucus and intestinal contents of H. otakii in facilities farm were extracted, the V3+V4 gene of different samples was analyzed by 16S rRNA high-throughput sequencing technology and biological analysis method. Results showed that there were only 33 common OTUs among different samples of skin mucus and intestinal contents. The Venn showed some similarities and differences among different samples. The rank abundance curve showed the homogeneity and richness of the samples, and these data were reasonable and reliable. The mean value of alpha index showed that the microbial richness of intestinal contents was high, and the species diversity of skin mucus was high; beta diversity showed the heterogeneity of different samples and the similarity of the same samples. At the phylum level, the dominant phylums were Cyanobacteria, Proteobacteria and Firmicutes. At the genus level, the core flora in intestinal contents included Streptophyta, Bacillus, Bacillaceae_unclassified, Aeromonas and Vibrio; while the core flora in skin mucus included Streptophyta, Aeromonas, Bacillaceae_unclassified, Myriudes, Gemmobacter and Vibrio. The heatmap showed the differences in the microbial species were biological functions, and the existence of conditional pathogens such as Bacillaceae_unclassified, Aeromonas, Vibrio, Gemmobacter might lead to the occurrence of diseases. In this study, the diversity and abundance of microorganisms and the characteristics of the main pathogenic bacteria in the skin mucus and intestinal contents were analyzed, which has an important significance for the healthy aquaculture and disease control of H. otakii.
    • In vitro Interaction Between Nitrogen Regulatory Protein GlnK with the Carbon Signaling Molecule α-ketoglutarate in Nitrogen-fixing Pseudomonas stutzeri
    • WANG Shanshan, WU Shaoyu, LIU Yichao, ZHAN Yuhua, KE Xiubin, LU Wei, YAN Yongliang*
    • 2021, 11(1):  91-98.  DOI: 10.19586/j.2095-2341.2020.0083
    • Abstract ( ) PDF (7061KB)( )
      • PⅡ protein is a general regulator of nitrogen metabolism in bacteria, which can sense the changes of carbon and nitrogen signals in cells and fine-tune the interaction between itself and the target proteins, so as to achieve the purpose of cascade regulation of nitrogen fixation. α-ketoglutarate is an important signal molecule of intracellular carbon status. Previous studies have found that PⅡ protein can bind α-ketoglutarate to sense intracellular carbon status, but there are differences in different bacteria. Pseudomonas stutzeri A1501 contains only one PⅡ protein GlnK, which plays an important role in the regulation of nitrogen fixation in A1501, but the specific carbon signal perception and transduction mechanisms needs to be further explored. Based on this, GlnK protein was expressed in Escherichia coli exogenously, and the in vitro interaction between GlnK protein and carbon signal molecule α-ketoglutarate was studied by microscale thermophoresis (MST) method. It was found that GlnK could interact with the signal molecule α-ketoglutarate in vitro, and it was further confirmed that the 89th glycine (G89, Gly89) of GlnK protein might be related to the interaction. The results laid a foundation for further analysis of α-ketoglutarate signal transduction in A1501, and also provided theoretical support for the in-depth analysis of the coupling mechanism of carbon and nitrogen metabolism of nitrogen-fixing bacteria.
    • Cloning of Series Truncated Gene Mutants by Partial Amplification Combining with Double Fragment Ligation
    • LIANG Mingxing, MA Mengqi, CHENG Yingying, WANG Yuan, CHEN Huabo*
    • 2021, 11(1):  111-117.  DOI: 10.19586/j.2095-2341.2020.0076
    • Abstract ( ) PDF (7486KB)( )
      • The random mutation associated with DNA synthesized in vitro is an important factor that restricts the efficiency of site-directed mutation. Taking cyclin E and its truncated genes as an example, in order to clone their kinase-deficient mutants, the traditional overlap extension PCR (OE-PCR) was improved appropriately, and an improved scheme about site-directed mutagenesis for series related genes was proposed. In the previous study, cyclin E gene and its two truncated genes T1 and T2 were cloned and inserted into pEGFP_C2 vector by EcoR Ⅰ/Sal Ⅰ double digestion. Restriction analysis showed that cyclin E gene sequence contained one Age Ⅰ site, which was divided into F1 and F2 segments, and the target mutation site KD was located in F2 segment. The F2 segment was identical to cyclin E and its truncated gene. Therefore, the consensus fragment F2 containing target mutation was amplified by overlap extension PCR (OE-PCR), and the corresponding F1 fragments were cut out from the three original plasmids C2-cyclin E, C2-cyclin E_T1 and C2-cyclin E_T2, and then F1 and the enzyme digested F2 were ligated into the vector to reconstruct the complete mutant. Comparative detection showed that OE-PCR was more successful in amplifying shorter DNA fragments. A certain number of clones were screened from C2-cyclin E, C2-cyclin E_T1 and C2-cyclin E_T2. After detection and DNA sequencing, one target mutant with completely accurate sequence was obtained from each group. Studies have shown that partial amplification could shorten the length of DNA synthesis, avoid the unfavorable factors such as repeated amplification of target genes, and thus reduce random mutations. And double-fragment ligation avoided the restriction of double Age Ⅰ sites on routine restriction-ligation. The combination of the two methods could be used as an optimization scheme for site-directed mutation of other related genes.

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