Abstract:In this paper, calmodulin gene of the HAB species Alexandrium catenella was isolated, and its transcription profile with respect to grow rate and heat stress was investigated. A full cDNA sequence of cam gene was obtained. It consisted of 597 nucleotides (nt), comprising 25 nt of the 5′ untranslated region (UTR), 122 nt of the 3′ UTR, and an open reading frame (ORF) of 450 nt, encoding 149 amino acid residues. The deduced CaM was highly conserved compared with CaM of other organisms. By implementing quantitative PCR, the relationship between transcription level of cam and growth rate of Alexandrium catenella was investigated. The results showed that cam expression level had a similar trend with the cell growth rate through the whole growing stage. The cam transcription was increased more than 8 fold in abundance from lag phase to the exponential phase, and obviously decreased from exponential phase to stationary/decline phase. Additionally, response of cam gene to heat stress was studied. Relative expression level of cam exhibited a significant decline trend with time during the heat shock experiment. All the results suggested that cam play an important role within process that the cells response to environmental stress and growth.

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	1. Cloning, characterization and expression analysis of calmodulin gene from Alexandrium catenella (Dinoflagellate) with respect to cell growth and heat stress
	WEN Ruobing,SUI Zhenghong
	Biology 01 June 2011
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	Abstract:In this paper, calmodulin gene of the HAB species Alexandrium catenella was isolated, and its transcription profile with respect to grow rate and heat stress was investigated. A full cDNA sequence of cam gene was obtained. It consisted of 597 nucleotides (nt), comprising 25 nt of the 5′ untranslated region (UTR), 122 nt of the 3′ UTR, and an open reading frame (ORF) of 450 nt, encoding 149 amino acid residues. The deduced CaM was highly conserved compared with CaM of other organisms. By implementing quantitative PCR, the relationship between transcription level of cam and growth rate of Alexandrium catenella was investigated. The results showed that cam expression level had a similar trend with the cell growth rate through the whole growing stage. The cam transcription was increased more than 8 fold in abundance from lag phase to the exponential phase, and obviously decreased from exponential phase to stationary/decline phase. Additionally, response of cam gene to heat stress was studied. Relative expression level of cam exhibited a significant decline trend with time during the heat shock experiment. All the results suggested that cam play an important role within process that the cells response to environmental stress and growth.
	TO cite this article:WEN Ruobing,SUI Zhenghong. Cloning, characterization and expression analysis of calmodulin gene from Alexandrium catenella (Dinoflagellate) with respect to cell growth and heat stress[OL].[ 1 June 2011] http://en.paper.edu.cn/en_releasepaper/content/4430812


	2. The complete genome and gene structure of mitochondrial DNA of African slender-snouted crocodile (Crocodylus cataphractus)
	Zhang Man ,Li Yan,Wang Yishu ,Wu Xiaobing
	Biology 17 March 2010
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	Abstract:The complete mitochondrial genome (mtDNA) of the African slender-snouted crocodile (Crocodylus cataphractus) was sequenced in this article. The whole length of the genome is 16,847 base pairs (bp). Just like other vertebrates, it has codes for 22 tRNAs, 13 protein-coding genes, 2 rRNAs, as well as a control region (D-loop). The gene order and the rearrangement of tRNA genes (tRNAPhe and tRNASer(AGY)) conform to that of other crocodilians sequenced, differing from typical vertebrates structure. It shows again that the gene order of crocodilians is considerably conserved.
	TO cite this article:Zhang Man ,Li Yan,Wang Yishu , et al. The complete genome and gene structure of mitochondrial DNA of African slender-snouted crocodile (Crocodylus cataphractus)[OL].[17 March 2010] http://en.paper.edu.cn/en_releasepaper/content/40799

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	3. Soluble expression and two-step purification of the cytoplasmic domains of Arabidopsis CORYNE in Escherichia coli
	Qu Wei,Zhao heng,Li ShuZhen,Luo Yunbo
	Biology 02 March 2010
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	Abstract:CORYNE (CRN) is a receptor-like protein kinase involved in control of Arabidopsis stem cell proliferation with CLAVATA2. To produce large amounts of soluble cytoplasmic kinase domains of CRN from Escherichia coli, three vectors were constructed using a double tag strategy, named pGEX-CRN, pMAL-CRN and p30-CRN. An auto-induction protocol was used to enhance the overproduction of soluble recombinant CRN with a 32Y media at 28 °C. The production of soluble recombined CRN of these vectors was analyzed and the pGEX-CRN was chosen for large-scale protein production. The dual-tagged GST-CRN-His6 was purified in a two-step affinity chromatography and analyzed for autokinase activity.
	TO cite this article:Qu Wei,Zhao heng,Li ShuZhen, et al. Soluble expression and two-step purification of the cytoplasmic domains of Arabidopsis CORYNE in Escherichia coli[OL].[ 2 March 2010] http://en.paper.edu.cn/en_releasepaper/content/40350


	4. 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


	5. 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


	6. 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


	7. Isolation of Trichoderma reesei PyrG negative mutant by UV Mutagenesis and its Application in Transformation
	Long Hao,Wang Tianhong,Zhang Yingkuan
	Biology 17 December 2007
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	Abstract:Two uridine auxotrophic mutants of Trichoderma reesei were isolated by resistance to 5-fluoroorotic acid after UV mutagenesis. One mutant, called M23, was complemented with the Aspergillus niger pyrG gene carried by plasmid pAB4-1. Mutated pyrG gene of M23 was cloned and DNA sequencing analysis indicated that a cytosine inserted into the 934-939 oligo dC position of the pyrG coding region, resulting in a frameshift mutation. Transformation efficiency was approximately 200-300 transformants per μg of DNA with plasmid pAB4-1. Stable transformants were obtained by monosporic culture and showed to be prototroph after successive propagation. Vitreoscilla hemoglobin expression plasmid pUCVHb co-transformed with plasmid pAB4-1 attained transformation efficiency of 71.8% versus 21.6% with pAN7-1. Southern blot analysis of the transformants demonstrated that plasmid pUCVHb was integrated into the chromosomal DNA. The experimental results demonstrated that the pyrG-based system was more efficient and timesaving than the conventional hygromycin B resistance-based transformation system.
	TO cite this article:Long Hao,Wang Tianhong,Zhang Yingkuan. Isolation of Trichoderma reesei PyrG negative mutant by UV Mutagenesis and its Application in Transformation[OL].[17 December 2007] http://en.paper.edu.cn/en_releasepaper/content/17054


	8. Effect of Calnexin Deletion on the Expession Level of Binding Protein (BiP) Under Heat Stress Conditions in Saccharomyces cerevisiae
	Zhang Huili ,Hu Bingjie ,Ji Yanyan ,Akio Kato,Song Youtao
	Biology 14 December 2007
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	Abstract:In order to investigate the effect of calnexin deletion on the induction of main ER molecular chaperone BiP, we cultured the wild-type and calnexin-disrupted Saccharomyces cerevisiae strains under the normal and stress conditions. The growth rate of the calnexin-disrupted yeast was almost the same as that of the wild-type yeast under those conditions. However, the induced mRNA level of BiP in the ER was evidently higher in calnexin-disrupted S. cerevisiae relative to the wild type at 37 C, but was almost the same in the two strains under the normal conditions. The western blotting analysis for BiP protein expression in the ER yielded results that showed a parallel in their mRNA levels in both strains. It is suggested that under heat stress conditions the induction of BiP in the ER could recover parts of the function of calnexin in calnexin-disrupted yeast, and result in the same growth rate as in wild-type yeast.
	TO cite this article:Zhang Huili ,Hu Bingjie ,Ji Yanyan , et al. Effect of Calnexin Deletion on the Expession Level of Binding Protein (BiP) Under Heat Stress Conditions in Saccharomyces cerevisiae[OL].[14 December 2007] http://en.paper.edu.cn/en_releasepaper/content/16921


	9. Cloning, Expression and Characterization of an Enantioselective Lipase LipB68 from Pseudomonas Fluorescens Strain B68
	Luo Yu ,Zheng Yitao,Jiang Zhengbing,Ma Yushu,Wei Dongzhi
	Biology 02 March 2006
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	Abstract:A lipase-producing bacterium strain B68 was screened and identified as Pseudomonas fluorescens sp. Novel lipase gene lipB68 was cloned and sequenced… by the method of GenomeWalker. Sequence analysis revealed an open reading frame of 1,431 nucleotides, encoding a protein with 476 amino acids and a molecular mass of 50,193 Da, being a member of lipase subfamily I.3. The lipB68 gene was expressed in E. coli BL21 (DE3) in the form of inclusion body and the purification and protein refolding were conducted. Lipase lipB68 had a favorable reaction temperature of 20°C with p-Nitrophenyl caprate as substrate, lower than all the known lipases from subfamily I.3. And it also showed good stability to organic solvents such as DMF and acetonitrile. After being immobilized on fabric of cellulose, lipase lipB68 showed high enantioselectivity at the resolution of racemic a-phenylethanol and a-phenylpropanol (E=220, and 190 respectively).
	TO cite this article:Luo Yu ,Zheng Yitao,Jiang Zhengbing, et al. Cloning, Expression and Characterization of an Enantioselective Lipase LipB68 from Pseudomonas Fluorescens Strain B68[OL].[ 2 March 2006] http://en.paper.edu.cn/en_releasepaper/content/5463

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