• Title/Summary/Keyword: Protein-1

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Functional Screening for Cell Death Suppressors and Development of Multiple Stress-Tolerant Plants

  • Moon Hae-Jeong;Baek Dong-Won;Lee Ji-Young;Nam Jae-Sung;Yun Dae-Jin
    • Journal of Plant Biotechnology
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    • v.5 no.3
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    • pp.143-148
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    • 2003
  • Bax, a mammalian pro-apoptotic member of the Bcl-2 family induces cell death when expressed in yeast. To investigate whether Bax expression can induce cell death in plant, we produced transgenic Arabidopsis plants that contained murine Bax cDNA under control of a glucocorticoid-inducible promoter. Transgenic plants treated with dexamethasone, a strong synthetic glucocorticoid, induced Bax accumulation and cell death, suggesting that some elements of cell death mechanism by Bax may be conserved among various organisms. Therefore, we developed novel yeast genetic system, and cloned several Plant Bax Inhibitors (PBIs). Here, we report the function of two PBIs in detail. PBI1 is ascorbate peroxidase (sAPX). Fluorescence method of dihydrorhodamine123 oxidation revealed that expression of Bax in yeast cells generated reactive oxygen species (ROS), and which was greatly reduced by co-expression with sAPX. These results suggest that sAPX inhibits the generation of ROS by Bax, which in turn suppresses Baxinduced cell death in yeast. PBI2 encodes nucleoside diphosphate kinase (NDPK). ROS stress strongly induces the expression of the NDPK2 gene in Arabidopsis thaliana (AtNDPK2). Transgenic plants overexpressing AtNDPK2 have lower levels of ROS than wildtype plants. Mutants lacking AtNDPK2 had higher levels of ROS than wildtype. $H_2O_2$ treatment induced the phosphorylation of two endogenous proteins whose molecular weights suggested they are AtMPK3 and AtMPK6. In the absence of $H_2O_2$ treatment, phosphorylation of these proteins was slightly elevated in plants overexpressing AtNDPK2 but markedly decreased in the AtNDPK2 deletion mutant. Yeast two-hybrid and in vitro protein pull-down assays revealed that AtNDPK2 specifically interacts with AtMPK3 and AtMPK6. Furthermore, AtNDPK2 also enhances the MSP phosphorylation activity of AtMPK3 in vitro. Finally, constitutive overexpression of AtNDPK2 in Arabidopsis plants conferred an enhanced tolerance to multiple environmental stresses that elicit ROS accumulation in situ. Thus, AtNDPK2 appears to playa novel regulatory role in $H_2O_2$-mediated MAPK signaling in plants.

The Strategy for the Development of Bio-Resources Utilizing Sericultural Products and Insects

  • Lee, Won-Chu;Kim, Iksoo
    • International Journal of Industrial Entomology and Biomaterials
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    • v.1 no.2
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    • pp.95-102
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    • 2000
  • Experiments related to the field of sericulture started in the years 1900, in Korea. The sericultural experimental station in Korea was first organized among agricultural fields in Korea, indicating that sericulture in Korea was regarded as an important field of agriculture. Sericulture has been devoted to a great deal for the improvement of Korean economy during the past 100 years even under the coarse social circumstances caused particularly by the Korean War, However, the traditional Korean sericulture, aimed to produce silk yarn, was weakened, because of several reasons such as diminishment in silk consumption, increased labor charge in Korea, and so on. After this difficulty time, the Korean sericulture was revolutionized by shifting into functional sericulture from 1995, and the Korean sericulture now plays an important role for the improvement of human health. Mulberry tree, silkworm, and silk have a boundless potential to be developed as resources. We expect the know-how obtained through silkworm research would expand to the other insect research too. Thus, an area of entomological industry is hoped to prosper owing to insect research as well as sericulture. Mulberry tree is known to possess many bio-active substances, so it can be utilized as a resource for substitute medicine and a raw material for the functional food. In addition, an invention of genetically engineered mulberry variety, which will produce more bioactive substances, is expected. Silkworm is one of the most extensively studied insect organisms on the genome so far, Thus, silkworm is expected to be an "insect bio-factory", enabling mass-production of useful proteins by transformation, in which useful foreign genes are assimilated into silkworm. Silk can be transformed into several phases, because it possesses useful functional groups, which are sensitive to chemical reaction. Also, because silk fibrin itself is protein, it has a superior applicability as tissue membrane. Due to this usefulness, many researchers are now working on the silk as food, cosmetic, medical resource, and bioengineering resource, and even an expanded application is expected using silk in the future. Until now, the researches on insects were largely focused on the prevention of the damage caused by pest, instead of a beneficial aspect. However, insects are thought to be the fourth natural resource in the world, possessing unlimited potential as world resources in the near future. Therefore, our entomological research effort should be focused on the subject with potential for industrialization. Such subject includes selecting the insect species useful for environmental evaluation, construction of environment-friendly agricultural ecosystem, pollen mediation, pet, and advanced bio-resources.

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Study on the Origins and Main Treatments of Grub Used in Experiments, and Research Directions on the Efficacy of Grubs (실험에 사용된 제조의 기원과 주치증에 대한 고찰 및 제조의 약효에 대한 연구방향)

  • Han, Kyu-Jo;Kim, Myung-Dong
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.23 no.1
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    • pp.158-165
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    • 2009
  • The aim of this study was to clarify the origins of grubs which were used in the experiments, and to determine many other effects that are now written in the classical medical and herbal books. We compared the efficacy of grubs written in the herbal and medical books on the one hand, and studied the origins of grubs used in the experiments, on the other hand. The following results were obtained. There are three kinds of grub species for medicinal uses. Holorichia diomphalia Bates mainly produced in China, Cetoniidae species mainly produced in Korea, and Protaeria brevitarsis in Korean folk remedy are three species. Grubs were used in 25 experiments. Of these experiments, Holotrichia diomphalia were used 12 times, Larve of Protaeria brevitarsis 3 times, and Protaetia orientalis was used once. So, of total 25 experiments, 36%(9 experiments) did not clarify the origins of grubs used in the experiments. Currently, the grubs, which are imported from China and used in Korea, are Holotrichia diomphalia Bates from China. Current experimental study shows that grubs have efficacy for protecting liver. So the use of grubs in folk medicine is believed to be based on the scientific evidence. Efficacy of grubs, though it is not recorded in the medical books, has the effects of anti-cancer(anti-tumor), antioxidant, and anti-diabetes mellitus. The ingredients of Holotrichia are protein, peptide, fat, enzyme, mineral, and other several nutrients. Study on the grubs so far has a tendency to be done with little focus on the differen of origin of grubs. So, from now on, it needs that the origin of grubs be clarified, and the study on the efficacy from each origin be done. Considering the effects that the classical herbal books have recorded, it is necessary that more profound study proceed to prevent and treat diseases on eyes and ears, on the basis of the fact that grub has multiple ingredients to replenish the vital essence.

Influence of Coated Urea Complex Fertilizer Application on Growth and Grain Quality of Paddy Rice (피복요소복합비료(被覆尿素複合肥料)의 시용방법(施用方法)이 벼 생육(生育)과 미질(米質)에 미치는 영향(影響))

  • Park, Kyeong-Bae
    • Korean Journal of Soil Science and Fertilizer
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    • v.26 no.2
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    • pp.72-77
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    • 1993
  • This study was conducted to evaluate coated urea complex fertilizer (CUC) application for rice growth and its grain quality on clay loam (Deogpyeong series) and sandy loam (Gangseo series) in southern region of Korea, 1992. The coated urea complex fertilizer used in the experiment was CUC I and II. The CUC I is a complex fertilizer consisting of 18-12-13 for $N-P_2O_5-K_2O$ including 50% of coated urea as nitrogen source, and CUC II is the same complex fertilize as mentioned above but it contains 25% of coated urea. Seventy percent of conventional fertilizer for nitrogen level was applied as basal dose of side-band placement (SBP) and whole plowlayer placement (WPP). Greenish degree of rice leaf color at the late growth stage was high in CUC plots regardless of application method and soil series. Amount of nitrogen uptaken by the rice plant at harvesting stage was mush more in the CUC treatment compared to convential fertilizer, and also much more in CUC I than that of CUC II. In the WPP on clay loam, harvest index of CUC treatment was showed very low level, which was reduced more in CUC II plot. In the CUC treatments, perfect rice grain ratio decreased remarkably, and the notched-belly rice kernel ratios among imperfect rice grain increased drastically. In rice grain, Hon-value(Mg to K.N ratio) decreased, protein and amylose contents increased in CUC application plots in WPP. In addition, gel consitency and alkali spreading value related to eating quality in CUC plots were getting longer and lower, respectively. The yield in CUC plot was similar to conventional fertilizing, but significant decreased in SBP. As a result, the rice grain quality became poor with the coated urea complex fertilizer application, but it could be improved by incorporation with fast-release fertilizer.

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Effects of Linseed Oil or Whole Linseed Supplementation on Performance and Milk Fatty Acid Composition of Lactating Dairy Cows

  • Suksombat, Wisitiporn;Thanh, Lam Phuoc;Meeprom, Chayapol;Mirattanaphrai, Rattakorn
    • Asian-Australasian Journal of Animal Sciences
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    • v.27 no.7
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    • pp.951-959
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    • 2014
  • The objective of this study was to determine the effects of linseed oil or whole linseed supplementation on performance and milk fatty acid composition of lactating dairy cows. Thirty six Holstein Friesian crossbred lactating dairy cows were blocked by milking days first and then stratified random balanced for milk yields and body weight into three groups of 12 cows each. The treatments consisted of basal ration (53:47; forage:concentrate ratio, on a dry matter [DM] basis, respectively) supplemented with 300 g/d of palm oil as a positive control diet (PO), or supplemented with 300 g/d of linseed oil (LSO), or supplemented with 688 g/d of top-dressed whole linseed (WLS). All cows were received ad libitum grass silage and individually fed according to the treatments. The experiment lasted for 10 weeks including the first 2 weeks as the adjustment period, followed by 8 weeks of measurement period. The results showed that LSO and WLS supplementation had no effects on total dry matter intake, milk yield, milk composition, and live weight change; however, the animals fed WLS had higher crude protein (CP) intake than those fed PO and LSO (p<0.05). To compare with the control diet, dairy cow's diets supplemented with LSO and WLS significantly increased milk concentrations of cis-9,trans-11-conjugated linoleic acid (CLA) (p<0.05) and n-3 fatty acids (FA) (p<0.01), particularly, cis-9,12,15-C18:3, C20:5n-3 and C22:6n-3. Supplementing LSO and WLS induced a reduction of medium chain FA, especially, C12:0-C16:0 FA (p<0.05) while increasing the concentration of milk unsaturated fatty acids (UFA) (p<0.05). Milk FA proportions of n-3 FA remarkably increased whereas the ratio of n-6 to n-3 decreased in the cows supplemented with WLS as compared with those fed the control diet and LSO (p<0.01). In conclusion, supplementing dairy cows' diet based on grass silage with WLS had no effect on milk yield and milk composition; however, trans-9-C18:1, cis-9,trans-11-CLA, n-3 FA and UFA were increased while saturated FA were decreased by WLS supplementation. Therefore, it is recommended that the addition 300 g/d of oil from whole linseed should be used to lactating dairy cows' diets.

Effect of Non-starch Polysaccharides and Resistant Starch on Mucin Secretion and Endogenous Amino Acid Losses in Pigs

  • Morel, Patrick C.H.;Melai, J.;Eady, S.L.;Coles, G.D.
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.11
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    • pp.1634-1641
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    • 2005
  • Generally, dietary fibre (DF) includes lignin, non-starch polysaccharides (NSP) and resistant starch (RS). In monogastric species, low levels of dietary fibre in the diet are associated with various diseases and high levels reduce nutrient digestibilities. In this study, the effects of different types and levels of NSP (soluble: $\beta$-glucan, insoluble cellulose) and resistant starch on mucin secretion and endogenous nitrogen and amino acid losses in pigs were investigated. A total of 25 five-week-old weaner pigs (9.5 kg${\pm}$1.5 kg), were randomly allocated to each of five experimental diets. Different levels of purified barley $\beta$-glucan (BG) extract (5 or 10% of $Glucagel^{(R)}$ $\beta$-glucan, providing 4 or 8% of BG in the diet), and resistant starch (RS) (8.3 or 16.6% of Hi-$Maize^{TM}$, providing 5 or 10% RS in the diet) were substituted for wheat starch in a purified diet in which enzymatically-hydrolysed casein was the sole source of protein. The diets were fed for 21 days. No statistically significant difference between treatments (p>0.05) was observed for growth performance and organs weights. No difference in ileal starch digestibility was observed between pigs on the cellulose or $\beta$-glucan diets. However, as the level of resistant starch in the diet increased the ileal starch digestibility decreased (p<0.05). The inclusion of resistant starch in the diet (5 or 10%) did not increase mucin production when compared with the cellulose-only diet. However, as the level of beta-glucan in the diet increased, both crude mucin in the digesta dry matter and per kg dry matter intake increased (p<0.05). Pigs fed the diet containing 8% of beta-glucan had higher endogenous loss flow than those fed the diets including 5 or 10% of resistant starch or 4% of $\beta$-glucan. In conclusion, dietary inclusion of resistant starch increased the level of starch reaching the large intestine without any effect on mucin secretion, or endogenous nitrogen or amino acid losses content in the small intestine. The addition of $\beta$-glucan to a diet containing cellulose increases both mucin secretion and endogenous amino acid and nitrogen losses in the small intestine.

Growth, Feed Efficiency, Behaviour, Carcass Characteristics and Meat Quality of Goats Fed Fermented Bagasse Feed

  • Ramli, M.N.;Higashi, M.;Imura, Y.;Takayama, K.;Nakanishi, Y.
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.11
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    • pp.1594-1599
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    • 2005
  • The effects of long-term feeding of diets based on bermudagrass hay supplemented with lucerne hay cube (LH) or fermented bagasse feed (FBF) on the growth rate, feed efficiency, behaviour, gut development, carcass characteristics and meat quality of goats were investigated. Six spring-born 8-month-old male crossbred (Japanese Saanen${\times}$Tokara native goats) bucks weighing mean 21.6 kg were allotted to 2 treatment groups (3 animals each) and each animal had ad libitum access to feeds, i.e. bermudagrass hay (basal diet)+LH or FBF throughout the experiment. The FBF was produced by the solid-state fermentation of substrates containing dried sugarcane bagasse mixed with wheat bran in a ratio of 1:3 (w/w DM) with Aspergillus sojae. The live body weight, final weight and average daily gain were not different between treatments. Average basal diet intake of goats fed FBF diet was significantly higher than that fed LH diet (p<0.05), but average dry matter intake (DMI; g/day and g/$W^{0.75}$), feed conversion ratio, digestible crude protein (DCP) and total digestible nutrients (TDN) intake of experimental diets were not significantly different between treatments. Goats fed on LH and FBF diets had similar eating, rumination, resting and drinking behaviours, and blood constituents except for phosphorus content. Slaughter and carcass weights, net meat percentage [(total meat/carcass weight)${\times}$100], loin ratio [(loin/total meat)${\times}$100] and rib-eye area were not different between treatments. However, goats fed FBF diet had lower dressing percentage and higher bone/muscle ratio compared with goats fed LH diet (p<0.01). Empty gut and guts fill of goats fed FBF diet were significantly greater (p<0.05 and p<0.01, respectively) than those fed LH diet. The weights of rumen and abomasum were also significantly heavier in goats fed FBF diet (p<0.05), but the length and density of papillae of rumen in goats were not different between treatments. Although meat composition of loin was not different in both groups, the meat of goats fed FBF diet was superior to that of LH diet in flavor, aroma and overall quality of loin (p<0.01). In conclusion, the nature of the diet consumed voluntarily did not affect subsequent growth, nutrient intake and behaviour of goats but had an influence on carcass traits and sensory evaluation of meat partly, when either of LH or FBF was fed with bermudagrass hay.

Use of N-alkanes to Estimate Intake and Digestibility by Beef Steers

  • Premaratne, S.;Fontenot, J.P.;Shanklin, R.K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.11
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    • pp.1564-1568
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    • 2005
  • The objective of the study was to evaluate the use of n-alkanes to estimate DM intake and digestibility by beef cattle. Six steers were blocked (3 blocks, 2 animals/block) according to the body weight (279${\pm}$19 kg) and randomly allotted within blocks to two diets (3 steers/diet). A second trial was conducted with the same animals (321${\pm}$18 kg) after 36 days (d), using a switch back design. The diets consisted of two types of chopped sun-cured hay, alfalfa (Medicago sativa L) hay, or fescue (Festuca arundinacea Schreb) and alfalfa mixture, which were fed in equal amounts to steers. Animals were dosed with $C_{32}$ and $C_{36}$ alkanes, employing an intra-ruminal controlled-release device at the beginning of each trial. Hay intake per animal was measured from d 6 to 12 and sub samples were taken for chemical analysis. Rectal samples of feces were taken from each animal once/daily from d 8 to 14, freeze dried, and ground prior to alkane analysis. Alkanes were extracted from ground hay and feces. Feed intake was calculated from the dose rate of $C_{32}$ alkane and, the herbage and fecal concentrations of adjacent odd ($C_{33}$ or $C_{31}$) and even ($C_{32}$) chain length alkanes. Crude Protein, NDF, ADF, ash concentrations and In vitro dry matter digestibility (IVDMD) were 17.7, 42.2, 28.4, 7.9 and 71.7 for alfalfa, and 12.4, 56.5, 30.4, 6.9 and 69.1% for fescue/alfalfa mixture, respectively. For both diets, intake estimated from $C_{33}$:$C_{32}$ ratio was not different from the measured intake, but intake estimated from $C_{31}$:$C_{32}$ ratio was lower (p<0.05), than the measured intake for both diets. The average estimated forage intake from $C_{33}$:$C_{32}$ ratio was 4.86 and 0.69% below than the measured intake for alfalfa and, fescue/alfalfa mixed diets, respectively. The respective estimates with $C_{31}$:$C_{32}$ ratio were 9.59 and 11.33% below than the measured intake. According to these results, alkane $C_{33}$:$C_{32}$ ratio is better than alkane $C_{31}$:$C_{32}$ ratio for the estimation of intake by beef steers.

Dietary L-carnitine Influences Broiler Thigh Yield

  • Kidd, M.T.;Gilbert, J.;Corzo, A.;Page, C.;Virden, W.S.;Woodworth, J.C.
    • Asian-Australasian Journal of Animal Sciences
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    • v.22 no.5
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    • pp.681-685
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    • 2009
  • L-carnitine promotes mitochondrial ${\beta}$-oxidation of long chain fatty acids and their subsequent transport across the inner mitochondrial membrane. Although the role of L-carnitine in fatty acid metabolism has been extensively studied, its role in live performance and carcass responses of commercial broilers is less understood. The objective of this research was to determine if Lcarnitine fed at various levels in diets differing in CP and amino acids impacted on live performance and carcass characteristics of commercial broilers. Two floor pen experiments were conducted to assess the effect of dietary L-carnitine in grower diets. In Exp. 1, Ross${\times}$Hubbard Ultra Yield broilers were placed in 48 floor pens (12 birds/pen) and fed common diets to d 14. A two (0 or 50 ppm Lcarnitine) by three (173, 187, and 202 g/kg CP) factorial arrangement of treatments was employed from 15 to 35 d of age (8 replications/treatment). An interaction (p<0.05) in carcass yield indicated that increasing CP (187 g/kg) resulted in improved yield in the presence of L-carnitine. Increasing CP from 173 to 202 g/kg increased (p<0.05) BW gain and decreased (p<0.05) feed conversion and percentage abdominal fat. Feeding dietary L-carnitine increased back-half carcass yield which was attributable to an increase (p<0.05) in thigh, but not drumstick, yield relative to carcass. In Exp. 2, $Ross{\times}Ross$ 708 broilers were fed common diets until 29 d. From 30 to 42 d of age, birds were fed one of seven diets: i) 200 g/kg CP, 0 ppm L-carnitine; ii) 200 g/kg CP, 40 ppm L-carnitine; iii) 180 g/kg CP, 0 ppm L-carnitine; iv) 180 g/kg CP, 10 ppm L-carnitine; v) 180 g/kg CP, 20 ppm L-carnitine; vi) 180 g/kg CP, 30 ppm L-carnitine; and vii) 180 g/kg CP, 40 ppm L-carnitine (6 replications of 12 birds each). BW gain, feed conversion, mortality (30 to 42 d), and carcass traits (42 d) were measured on all birds by pen. There were no treatment differences (p<0.05). However, the addition of 40 ppm L-carnitine in the 200 g CP/kg diet increased (p = 0.06) thigh yields relative to BW in comparison to birds fed diets without L-carnitine, which was further confirmed via a contrast analysis (0 vs. 40 ppm L-carnitine in the 200 and 180 g CP/kg diets; p<0.05). These results indicated that dietary L-carnitine may heighten metabolism in dark meat of commercial broilers resulting in increased relative thigh tissue accretion without compromising breast accretion.

Molecular Cloning and Characterization of the Yew Gene Encoding Squalene Synthase from Taxus cuspidata

  • Huang, Zhuoshi;Jiang, Keji;Pi, Yan;Hou, Rong;Liao, Zhihua;Cao, Ying;Han, Xu;Wang, Qian;Sun, Xiaofen;Tang, Kexuan
    • BMB Reports
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    • v.40 no.5
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    • pp.625-635
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    • 2007
  • The enzyme squalene synthase (EC 2.5.1.21) catalyzes a reductive dimerization of two farnesyl diphosphate (FPP) molecules into squalene, a key precursor for the sterol and triterpene biosynthesis. A full-length cDNA encoding squalene synthase (designated as TcSqS) was isolated from Taxus cuspidata, a kind of important medicinal plants producing potent anti-cancer drug, taxol. The full-length cDNA of TcSqS was 1765 bp and contained a 1230 bp open reading frame (ORF) encoding a polypeptide of 409 amino acids. Bioinformatic analysis revealed that the deduced TcSqS protein had high similarity with other plant squalene synthases and a predicted crystal structure similar to other class I isoprenoid biosynthetic enzymes. Southern blot analysis revealed that there was one copy of TcSqS gene in the genome of T. cuspidata. Semi-quantitative RT-PCR analysis and northern blotting analysis showed that TcSqS expressed constitutively in all tested tissues, with the highest expression in roots. The promoter region of TcSqS was also isolated by genomic walking and analysis showed that several cis-acting elements were present in the promoter region. The results of treatment experiments by different signaling components including methyl-jasmonate, salicylic acid and gibberellin revealed that the TcSqS expression level of treated cells had a prominent diversity to that of control, which was consistent with the prediction results of TcSqS promoter region in the PlantCARE database.