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	1. A Novel Tetracycline-Suppressed Expression System Based on Single Lentiviral Vector
	Yang Chunyan ,Liu Jia,Wang Chunhong,Sun Yan,Xu Zenghui,Zhou Chengliang,Lei Qiugang,Wu Mengchao,Qian Qijun
	Biology 01 December 2009
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	Abstract:Tetracycline-regulated eukaryotic expression system is a useful tool in area of biological researches. However, the current system is characterized of separation of essential elements into two individual vectors, resulting in inconvenience in the process of application. Here, we successfully generated single-vector tetracycline-regulated expression system which is based on recombinant lentivirus. Our data confirmed the efficiency and reliability of this single-vector controllable expression system. We hope that our work would advance the application of tetracycline-regulated system in future.
	TO cite this article:Yang Chunyan ,Liu Jia,Wang Chunhong, et al. A Novel Tetracycline-Suppressed Expression System Based on Single Lentiviral Vector[OL].[ 1 December 2009] http://en.paper.edu.cn/en_releasepaper/content/37123


	2. Expression Analysis and Characteristics of Prohibitin Gene from Silkworm, Bombyx mori
	Zhang Xuefang,Zhuang Wenhua,Lv Zhengbing,Zhang Yaozhou
	Biology 24 November 2009
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	Abstract:Prohibitin (PHB) is an evolutionarily conserved ubiquitously expressed multifunctional protein. However, its mechanism of action is unknown as yet. To better characterize the prohibitin protein from silkworm pupae (BmPHB), a gene encoding a prohibitin protein was identified from a cDNA library of silkworm pupae, which has an ORF of 825 bp, encoding a predicted 274 amino acids. A His-tagged BmPHB fusion protein was expressed in Escherichia coli Rosetta (DE3) and purified with affinity and reversed-phase chromatographies. Purified rBmPHB was used to generate anti-BmPHB polyclonal antibody, which was used to determine the subcellular localization of BmPHB. Immunostaining indicated that prohibitin can be found in both nucleus and cytoplasm but is located primarily in cytoplasm. Western blot analyses indicated that, in the fifth instar larva, BmPHB was expressed descendingly in gonad, malpighian tubule, trachea, fatty body, intestine and head. However, none was detected in larva’s silk gland and epidermis. Additionally, BmPHB was expressed in the nascent egg, larva and pupa, but none was detected in the moth. Therefore, we hypothesized that BmPHB may play an important role on silkworm development. This is a first description of the prohibitin protein from silkworm pupae, B. mori.
	TO cite this article:Zhang Xuefang,Zhuang Wenhua,Lv Zhengbing, et al. Expression Analysis and Characteristics of Prohibitin Gene from Silkworm, Bombyx mori[OL].[24 November 2009] http://en.paper.edu.cn/en_releasepaper/content/36970

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	3. Expression, Purification and Characterization of Malate dehydrogenase from Thermus thermophilus HB27
	Liu Jun,Lv Zhengbing,Meng Fanguo,Zhang Yaozhou
	Biology 16 November 2009
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	Abstract:Malate dehydrogenase (MDH, EC 1.1.1.37) is an essential metabolic enzyme in tricarboxylic acid cycle. The gene of the malate dehydrogenase of Thermus thermophilus HB27 was cloned and expressed in E.coli. The 984 bp gene encoded a homodimer enzyme of 35.39 kDa per subunit. The enzyme was purified by heat purification and metal chelate affinity chromatography. The relative activity was measured in different grades of pH value and temperature. The results showed that the optimal pH value was approximately 11.5 and the optimal temperature was 80℃. Our results may provide useful information regarding the biochemical properties of the MDHs from thermophilic bacteria.
	TO cite this article:Liu Jun,Lv Zhengbing,Meng Fanguo, et al. Expression, Purification and Characterization of Malate dehydrogenase from Thermus thermophilus HB27[OL].[16 November 2009] http://en.paper.edu.cn/en_releasepaper/content/36725


	4. The subgenus Stegana (Steganina) (Diptera: Drosophilidae) from Malaysia, with descriptions of four new species
	Chen Xipeng ,Chen Hongwei
	Biology 04 November 2009
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	Abstract:Seven species of the subgenus Stegana (Steganina) were found from eastern Malaysia, including four new species: S. (S.) angulata sp. nov., S. (S.) calyptraea sp. nov., S. (S.) platyphylla sp. nov. and S. (S.) obscura sp. nov. A key to species unassigned to any species group from Malaysia is provided.
	TO cite this article:Chen Xipeng ,Chen Hongwei . The subgenus Stegana (Steganina) (Diptera: Drosophilidae) from Malaysia, with descriptions of four new species[OL].[ 4 November 2009] http://en.paper.edu.cn/en_releasepaper/content/36393


	5. Novel fluorescent proteins generated by de novo synthesis method and their characterization
	Sun Tingting,Yi Ke,Zhu Cong,Wang Jinchun,Gao Xiaolian,Guo Jiangfeng
	Biology 28 September 2009
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	Abstract:Up to now the method to get fluorescent proteins (FPs) variants is focused on random mutation and site-directed mutagenesis. Herein we can not forecast and get the special expected mutated site at our will for random mutation. Though site-directed solved this problem, its inherent limitation that it is difficult to get amount mutation at the same time and it is a money-spending and time-consuming method hindered its application. In this article we used a high throughput and relatively cheap technology- microfludic PicoArray method for the simultaneous de novo synthesis fluorescent proteins（FPs）DNA sequences and reassemble sequences successively to get massive variants of fluorescent proteins. The result showed that five FPs are unique and have properties different to the others FPs existed and within these five new FPs, three FPs with different hues are belong to red fluorescent proteins which emission wavelength are longer than 550 nm and the other two locate at green fluorescent protein region. The native gel combined with Mass spectrum showed that all the five FPs are monomer with molecular weight are between 25 kD and 35 kD and they have relative stronger fluorescence intensity comparing with former blue fluorescent proteins, so they will be potential tools in molecular and medical studying fields.
	TO cite this article:Sun Tingting,Yi Ke,Zhu Cong, et al. Novel fluorescent proteins generated by de novo synthesis method and their characterization[OL].[28 September 2009] http://en.paper.edu.cn/en_releasepaper/content/35543


	6. cDNA Cloning ,Pituitary Location and Extra-pituitary Expression of Pro-opiodmelanocortin (POMC) Gene in Rare Minnow (Gobiocypris rarus)
	Liu Xiaohong,Xie Biwen,Zhang Yaoguang,Wang Deshou,Wang Zhijian
	Biology 27 September 2009
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	Abstract:A cDNA encoding pro-opiodmelanocortin (POMC) was cloned from pituitary of the rare minnow ( Gobiocypris rarus), a small freshwater fish endemic to China ，by polymerase chain reaction(RT-PCR) and rapid amplification of cDNA ends (RACE). Data showed that, the predicted rare minnow POMC cDNA (rmPOMC) consisted of 844bp with a 666bp ORF coding for the following sequences flanked by proteolytic cleavage sites: signal peptide (SP, Met1–Ala28), N-terminal peptide (Gln29–His105), ACTH (Ser108–Met146), α-MSH ( Ser108–Gal121), CLIP (Pro126–Met146), β-LPH (Glu149–His221), γ-LPH (Glu149– Ser186), β-MSH (Asp 170–Ser186), β-endorphin (β-EP, Tyr189–Gln221). Sequence analysis showed no region homologous to γ-MSH/ joining peptide (a tetrapod pomc feature ) was found. Amino acid sequence shares a highest similarity with POMC-Ⅰand POMC-Ⅱof common carp (92.4%) according to homologous alignment. Pituitary and extra-pituitary expression were studied by RT-PCR and in situ hybridization. rmPOMC positive cells mainly located in rostral pars distails (RPD) and pars intermedia(PI). Otherwise, there were some rmPOMC positive cells detected in PPD too according to in situ hybridization. In the extra-pituitary tissues, positive signals were observed in brain, intestine, gonad, hepatopancreas, spleen and gill by RT-PCR analysis.
	TO cite this article:Liu Xiaohong,Xie Biwen,Zhang Yaoguang, et al. cDNA Cloning ,Pituitary Location and Extra-pituitary Expression of Pro-opiodmelanocortin (POMC) Gene in Rare Minnow (Gobiocypris rarus)[OL].[27 September 2009] http://en.paper.edu.cn/en_releasepaper/content/35503


	7. The conservative gene regulatory network in HK97 and λ phages
	Ai Duiyuan,Qi Yanjiao
	Biology 08 June 2009
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	Abstract:Some conservative gene regulatory elements are found in Enterobacteria phage HK97 through sequences alignment, which are very similar to λ phage. Meanwhile, the regulated genes such as c1-gene, cro-gene, Q-gene, also show homology with λ phage. All of these results suggest hk97’s gene regulatory network is as same as λ phage. It implies this regulatory network is crucial and universal to lambda group.
	TO cite this article:Ai Duiyuan,Qi Yanjiao. The conservative gene regulatory network in HK97 and λ phages[OL].[ 8 June 2009] http://en.paper.edu.cn/en_releasepaper/content/32942


	8. Comparison of biochemical characterizations of AChE in the larvae of three populations of Tribolium castaneum (Herbst): Implications of insecticide resistance
	Wei Dandan,Liu Guoying,Jiang Hongbo,Dou Wei,Wang Jinjun
	Biology 10 March 2009
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	Abstract:The toxicological and biochemical characteristics of acetylcholinesterase (AChE) in two field and one laboratory populations of Tribolium castaneum larvae were investigated. The two field populations HSTC-R (higher level resistance) and HYTC-R (lower level resistance) were both resistant to phosphine as well as organophosphorus insecticides such as dichlorvos and malathion. Compared with the laboratory population (ABTC-S), the activities per insect and specific activities of AChE in HSTC-R and HYTC-R were significantly lower. The apparent Michaelis–Menten constant value (Km) for acetylthiocholine iodide (ATChI) was obviously lower in ABTC-S than that in HSTC-R, indicating a higher affinity to the substrate ATChI in the laboratory population. The affinity for the substrate ATChI in HYTC-R and ABTC-S were not significantly different. The Vmax value of the HSTC-R was significantly greater compared to the Vmax for the ABTC-S suggesting a possible over expression of AChE in this field population. The inhibition study of AChE to insecticide exposure in vitro revealed that all the inhibitors possessed excellent effect. For the efficiencies of the tested inhibitors, the rank order from the most sensitive to the least was eserine, followed by malaoxon, paraoxon-ethyl, carbaryl, and demeton-S-methyl. Based on the I50s, AChE of the ABTC-S were more sensitive to all the inhibitors than those of HSTC-R. The statistical analysis of the bimolecular rate constants (ki) was consistent with the above situation. It is concluded that the insensitive AChEs were probably involved in the resistance of T. castaneum against insecticides observed in the field populations.
	TO cite this article:Wei Dandan,Liu Guoying,Jiang Hongbo, et al. Comparison of biochemical characterizations of AChE in the larvae of three populations of Tribolium castaneum (Herbst): Implications of insecticide resistance[OL].[10 March 2009] http://en.paper.edu.cn/en_releasepaper/content/30071


	9. The fragment containing CP gene of Cucumber mosaic virus determines the severe leaf distortion phenotype expression on N. tabacum
	Xu Gang,Wu Peng,Liao Qian-sheng,Chen Ji-shuang
	Biology 04 March 2009
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	Abstract:The Cb7 isolate of Cucumber mosaic virus (CMV) induces severe leaf distortion on Nicotiana tabacum in contrast to CMV-Fny that causes systemic mosaic in this host. It was found that N. tabacum plants infected with the reassortant, a combination of transcripts derived from cDNAs of CMV-Fny RNA1 and 2 (F1 and F2) and of CMV-Cb7 RNA3 (F12C3), displayed severe leaf distortion symptoms characteristic of the wild-type Cb7 infection, indicating that genomic RNA3 had a distinctive role in the determination of the symptom. An exchange of the fragment containing CP gene between Cb7 and Fny constructed two chimeric RNA3s: CFB, including CP gene of Fny, and FCB, including CP gene of Cb7. Each chimera mixed with F1 and F2 was inoculated onto N. tabacum. The F12CFB-infected plants expressed systemic mosaic symptoms, while the F12FCB-infected ones showed severe leaf distortion symptoms. The results suggested that the genetic determinant of the severe leaf distortion symptom showing on N. tabacum was exclusively associated with the fragment containing CP gene in the Cb7 genome. Accumulation of CMV RNAs in the systemic leaves of N. tabacum plants infected with the reassortant was determined using Northern blotting. The results showed that the accumulation of subgenomic RNA4 of F12FCB was significantly higher than that of F12CFB and Fny, as a control, and the amounts of RNA4 of F12CFB were equivalent to those of Fny. Therefore, it had been inferred that the high amounts of viral subgenomic RNA4, presenting high accumulation of CP, might be responsible for the severe leaf distortion symptom expression on N. tabacum.
	TO cite this article:Xu Gang,Wu Peng,Liao Qian-sheng, et al. The fragment containing CP gene of Cucumber mosaic virus determines the severe leaf distortion phenotype expression on N. tabacum[OL].[ 4 March 2009] http://en.paper.edu.cn/en_releasepaper/content/29878


	10. Complete nucleotide sequences of two double-stranded RNAs from Raphanus sativus-root demonstrate the incidence of a novel cryptic virus
	Qinghua Tian,Liqing Li,Jishuang Chen
	Biology 26 February 2009
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	Abstract:Two double-stranded RNAs were extracted from leaf tissues of Raphanus sativus-root cv. Yidianhong grown in Eastern China. Their full sequences, namely RasR7 and RasR8, were determined by a single primer amplification technique. Comparisons of 5′ untranslated regions (UTR) and 3′ UTR between RasR7 and RasR8 showed that they were highly conserved at 5′ UTR and an identical region 5′-AGAAUUU-3′ was discovered as the evidence of the same plant virus. 5′-UAAGAC-3′ was found as their identical region in 3′ UTR. BLASTP search in Genbank showed that RasR7 encoded putative protein was much similar with RNA dependent RNA polymerases (RdRps) of plant-infecting dsRNA viruses belonging to the family Partitiviridae. Phylogenetic analysis showed that RasR7 encoded putative protein clustered with the RdRps of members of the family Partitiviridae. RT-PCR detection indicated that both RasR7 and RasR8 existed in the virus-like particles. Therefore, it was suggested that RasR7 and RasR8 institute the genome of a newly discovered cryptic virus, namely Raphanus sativus cryptic virus 3 (RasV3).
	TO cite this article:Qinghua Tian,Liqing Li,Jishuang Chen. Complete nucleotide sequences of two double-stranded RNAs from Raphanus sativus-root demonstrate the incidence of a novel cryptic virus [OL].[26 February 2009] http://en.paper.edu.cn/en_releasepaper/content/29713

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