Current Biotechnology

Current Situation, Challenge, Countermeasures and Suggestions of Industrial Biotechnology
TAN Tao1, ZHAO Li-ming2*
2016, 6(3): 153-158. DOI: 10.3969/j.issn.2095-2341.2016.03.01
Abstract ( ) PDF (1086KB)( )

The important status and functions of industrial biotechnology in the global sustainable development had been expatiated based on the development trends of international industrial biotechnology. The paper analyzed the development status and challenges of domestic industrial biotechnology, and proposed the countermeasures and suggestions for the development of domestic industrial biotechnology.

Development, Challenges and Countermeasures for Industrialization of Genetically Modified Staple Crops in China
DU Jian-zhong1, HAO Yao-shan1, WANG Yi-xue1, ZHANG Huan-huan1, SUN Yi1*, SHI Hong-wei2
2016, 6(3): 159-168. DOI: 10.3969/j.issn.2095-2341.2016.03.02
Abstract ( ) PDF (1168KB)( )

At present, only transgenic cotton and papaya are grown in China with large area. Although a few transgenic staple crops varieties, such as rice and maize, had been granted safety assessment certificate for nearly 7 years, they have not been used for commercial use now. In contrast to that, large amounts of maize and soybean products are imported to China every year, and most of them are genetically modified (GM) products. That is because, on the one hand that the current atmosphere of public opinions on GM crops and the research and development status of GM crops in our country require us to be prudent and cautious to advance transgenic staple crop commercialization; on the other hand, global development tendency, yield\|enhancing potential, economic and environmental benefits, and our countrys basic strategy to gain high ground in this agriculture high-tech field as well as the domestic market demand of GM crops commercial use require us to speed up the commercialization process of GM staple food crops. How to resolve this contradiction and to carry out transgenic staple food crop development and commercialization smoothly in China, it is now one of our main tasks at present. Therefore, it is necessary to discuss the current status, problems and countermeasures of commercial use of GM staple crops in China, hoping to provide reference for realizing GM staple food commercialization in our country in the near future.

Status on Pathogenic and Resistance Gene of Cucumber Target Leaf Spot
ZHANG Zi-xin1, XIE Xue-wen2, FU Jun-fan1*, LI Bao-ju2*
2016, 6(3): 169-173. DOI: 10.3969/j.issn.2095-2341.2016.03.03
Abstract ( ) PDF (1110KB)( )

Cucumber target leaf spot is a new epidemic disease of cucumber in china. Experiencing long-term scientific research, many breakthroughs had been achieved in the fields of pathogen and resistance genes. The research progress on the occurrence, characteristics, pathogenic and resistance gene of cucumber target leaf spot were summarized in this paper. The problems in present studies were pointed out. Moreover, the corresponding suggestion and research direction for future were discussed, in order to provide reference for the identification, control and resistance breeding of cucumber target leaf spot.

Progress on Application of Hydrogen in Cancer Prevention and Treatment
LI Jia-la, ZHANG Ya-ting, XIE Fei*
2016, 6(3): 174-178. DOI: 10.3969/j.issn.2095-2341.2016.03.04
Abstract ( ) PDF (1100KB)( )

As a novel antioxidant, molecular hydrogen exhibits great efficacy in various diseases caused by oxidative stress and inflammatory. With a complex pathogenesis, cancer is difficult to prevent and cure. Numbers of malignancies are associated with chronic inflammation and oxidative stress. Therefore, molecular hydrogen may play a potential role in carcinogenesis and cancer therapy. Previous studies showed that molecular hydrogen may have the preventive and therapeutic effect in various tumors, as well as the synergistic effect in the tumor radiotherapy and chemotherapy with a decreasing side-effect. This paper reviewed the recent progress of molecular hydrogen in cancer prevention and treatment, and prospected the research direction in the future, which was expected to provide reference for hydrogen application in cancer prevention and treatment.

Progress on Application of Mesoporous Silica Nanoparticles in Controlled Drug Release and Targeted Drug Delivery of Cancer Chemotherapy
MU Jing-hai, YE Zhou, ZHANG Quan*, YIN Jian*
2016, 6(3): 179-184. DOI: 10.3969/j.issn.2095-2341.2016.03.05
Abstract ( ) PDF (3052KB)( )

Cancer chemotherapy normally produces systemic toxicity and undesired side effects on patients. In order to improve the chemotherapy, its desiderated to develop new type of nanomaterials for controlled drug release and targeted drug delivery. Mesoporous silica nanoparticles (MSNs) have attracted much attention for their potential use as drug delivery carriers. The unique properties of MSNs, such as high surface area, uniform mesoporous, tunable nanoparticle size and easy chemical functionalization, make them highly suitable as a therapeutic delivery vehicle. This article reviewed recent progress on controlled drug release and targeted drug delivery of MSNs, and prospected development as drug delivery carriers for cancer therapy.

Advances of Endophytic Fungi from Artemisia annus
LI Ling-ling
2016, 6(3): 185-187. DOI: 10.3969/j.issn.2095-2341.2016.03.06
Abstract ( ) PDF (1063KB)( )

Plant endophytic fungi can produce secondary metabolites which are similar to the host plants. It has become an important production source of active compounds and discovery way of new compounds. In order to explore the potential application value of endophytic fungi from Artemisia annus, recent advances on the biodiversity, bioactive and metabolites of endophytic fungi from Artemisia annus were summarized. Some recommendations for future research of endophytic fungi from Artemisia annus were put forward, which was expected to provide references for the resource exploitations of endophytic fungi from Artemisia annus.

Studies on Leaf Surface Microflora of Cigar-wrapper During Artificial Fermentation
DU Jia1, ZHANG Xiao-juan1, WU Gang2, ZHOU Rui-sheng3, CUI Yu-hui4, SHI Xiang-dong1*
2016, 6(3): 188-192. DOI: 10.3969/j.issn.2095-2341.2016.03.07
Abstract ( ) PDF (5943KB)( )

The cigar-wrapper was fermented for 42 days in Humidity Chamber, and the tobacco leaf surface microbe was separated, purified and identified. The study discussed the changes of cigar-wrapper artificial fermentation process of microbial flora. The results showed that: In cigar-wrapper artificial fermentation process, bacterial was the dominant flora, and mold accounted for a small proportion, and actinomycetes and yeasts were not found; All the bacterials were bacteria Bacillus, the number descending order were as follows: Bacillus megaterium> Bacillus subtilis> Bacillus cereus> Bacillus circulans> Bacillus mycoides> Bacillus stearothermophilus>Bacillus pumilus>Bacillus coagulans; The number of bacteria in the cigar-wrapper surface fermentation process showed a sharp downward trend in the first 24 d; Bacillus megaterium and Bacillus subtilis were the dominant species of cigar-wrapper artificial fermentation. Bacillus megaterium accounted for more than 50% and Bacillus subtilis accounted for about 19%.

Cloning, Expression and Sequence Analysis of Amylase Gene in the Silkworm, Bombyx mori
LV Yan-na1,2, CHEN Xiao-yan1,2, SHAO Li-ping1,2, LI Yi-nv2, HU Xiao-yuan2, ZHANG Zhi-fang2, YI Yong-
2016, 6(3): 193-199. DOI: 10.3969/j.issn.2095-2341.2016.03.08
Abstract ( ) PDF (2977KB)( )

Amylase is one of the important digestive enzymes in silkworm and known for hydrolyzing starch and glycogen. The mRNA expression level of the Bmamy7 and Bmamy5 in different tissues of 5th3d larva was analyzed by real- time PCR. Absolute quantitative analysis results indicated, Bmamy7 expressed in midgut was up to 3×109 copies/μg total RNA, its expression level in the malpighian was low, only 1.9×106 copies/μg total RNA; however, Bmamy5 was only expressed in midgut and the mRNA expression level was 1.5×109 copies/μg total RNA. The specific primers were designed according to the cDNA sequence predicted encoding amylase in silkworm transcript database, two amylase genes were cloned by RT-PCR. The complete sequence of Bmamy7 is 1 608 bp, ORF consists 748 bp, encodes 503aa. Amino acid sequence analysis showed that Bmamy7 had a typical structure domain of α-amylase. The complete sequence of Bmamy5 is 1 196 bp，ORF consists 738 bp and encodes 245aa. Bmamy5 and Bmamy7 gene was expressed in prokaryotic expression respectively. The activity of recombinant Bmamy7 protein was very weak. The expression product of Bmamy5 was not detected by SDS-PAGE. The results provided references for research and application of amylase gene in the silkworm.

Rat Gastrointestinal Histopathological Changes and its Intestinal Flora Profile Analysis Using ERIC\|PCR after Acute Stress
ZHU Xiao-hui1,2, ZHANG Cheng-gang2, LIU Hai-feng 1,2*
2016, 6(3): 200-205. DOI: 10.3969/j.issn.2095-2341.2016.03.09
Abstract ( ) PDF (6097KB)( )

In order to study the feasibility of establishing the acute stress model based on water-immersion and restraint stress, and observe the histopathological change of stomach, ileum, cecum and colon using HE staining, and further identify the change rule of intestinal flora profile by ERIC-PCR. The total healthy male rats were randomly devided into two groups: control group and model group. Rats were regularly raised in control group, and model group were placed in a homemade constraint cages after inhalation anesthesia with isoflurane, and then immersed in 23±2℃ water for 5h when it revived. Then, the rat body weight was recorded and had on open field test and forced swimming test. Finally, rats were dislocated to collect the tissues and its contents of stomach, ileum, cecum and colon to perform HE staining and ERIC-PCR identifying respectively. We gathered the images to study the gastrointestinal histopathological changes and its intestinal flora profile analysis using ERIC-PCR after acute stress. Comparing with control, the rat body weight had no significantly variation, the number of through significantly decreased, the number of upright significantly decreased, the number of grooming significantly decreased, and the no moving time of forced swimming significantly prolonged. Furthermore, the gastric mucosa and the microvilli of intestinal tract were not damaged, and no nuclei deeply staining, and several obviously changing characteristic bands in the intestinal flora were appeared. This study successfully established the acute stress model, and followed with HE staining and ERIC-PCR to provide a significant reference of studying on the intestinal flora profile based on rat acute stress model, and had a significant application value.

Study on Ferrous Oxidase-encoding Genes of Acidithiobacillus ferrooxidans
ZHANG Peng-hua, LI Jiang*, XU Ling-ling
2016, 6(3): 206-211. DOI: 10.3969/j.issn.2095-2341.2016.03.10
Abstract ( ) PDF (2986KB)( )

Acidithiobacillus ferrooxidans is an important mineral-bioleaching bacterium with ferrous ion or reducible sulfide as the energy substance. Ferrous oxidase plays a key role in the ferrous ion oxidation process. In this study, four strains (YHC, 721, 737 and 745) were isolated from different mines showed more than 98% nucleotide sequence with the 16S rDNA of A. ferrooxidans. Ferrous oxidase activity assays indicated that strains YHC and 737 had similar activity, which were obviously higher than that of strains 721 and 745. Four ferrous oxidase-encoding genes from these Acidithiobacillus strains were amplified by PCR, sequenced and compared with reference sequences in GenBank. Three bases showed difference and accounted for the only divergence at position 73, Val for strains YHC and 737 and Ala for strains 721 and 745. When substituted Val with Ala by using the site-directed mutagenesis, the mutant enzymes showed decreased thermostability and shorter half-life. As results, the ferrous oxidase had reduced functional duration and less enzyme activity.

Study on the Preparation of Recombinant Human Interferon α2b
LIANG Guo-yi, LIU Xiao-hang
2016, 6(3): 212-218. DOI: 10.3969/j.issn.2095-2341.2016.03.11
Abstract ( ) PDF (2274KB)( )

Recombinant human interferon α2b was prepared by cloning, expression, purification for obtaining rhIFN α2b with high activity and purity. Overlap extension PCR was performed to obtain the gene encoding for IFN α2b. A prokaryotic vector expressing rhIFN α2b was constructed successfully. The expression vector pBV220-IFN α2b was transformed into E. coli strain DH5α. After fermentation, the bacteria cells was collected and lysed. Finally, the rhIFN α2b was purified by denaturation, renaturation, ion exchange chromatography and gel filtration chromatography, and its specific activity reached 1×108 IU/mg. The results was expected to lay the foundation for preclinical research and long-acting preparation.

Research and Appraisal on Purification Process of Recombinant Human Long-acting Follicle Stimulating Hormone
LIU Xiao-hang, GAO Li-ting
2016, 6(3): 219-223. DOI: 10.3969/j.issn.2095-2341.2016.03.12
Abstract ( ) PDF (2678KB)( )

To obtain high purity recombinant human long-acting follicle stimulating hormone FSH-CTP, the study collected the CHO cells culture supernatant contained FSH-CTP, then it was puried by dye affinity chromatography, DEAE anion exchange chromatography and hydrophobic interaction chromatography. We detected the puried samples by molecular weight, isoelectric point, biological activity in vitro (Western Blot) and biological activity in vivo (ovarian weight gain in female rats law). The results showed that: The purity of FSH-CTP protein could reach 98% after three stept purification, the apparent molecular weight was about 47 kDa, MALDI-TOF-MS result was 35.2 kDa. The results of isoelectric point was 4.0~2.8. And we provided the FSH-CTP activity was higher than recombinant human follicle stimulating hormone significantly by Western Blot analysis. The purification and activity of FSH-CTP was high. The results could lay the foundation for further study of the drug.

Effect of Harmonia axyridis Against Aphids on Cauliflower
JIAO Xue-xia1, JIA Qian1, YANG Jin-li1, WANG Wei-ping1, WANG Bu-yun2, GU Wei1, WANG Ya-nan1*
2016, 6(3): 224-226. DOI: 10.3969/j.issn.2095-2341.2016.03.13
Abstract ( ) PDF (1071KB)( )

To evaluate the control effect of Harmonia axyridis (Pallas) against aphids on broccoli, the study was carried out by releasing eggs of H. axyridis with 4 different ratio of beneficial to harmfulratios. The results showed that the control effect was best when beneficial to harmfulratio was 1∶10. Under this condition, the control effecacy reached 61.36% in 5 days, and was still at 59.23% after 28 days. The results indicated that the validity of Harmonia axyridis against aphids was better.

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