茚三酮比色法测定矿物表面吸附浸矿细菌蛋白质含量

为了研究浸矿细菌在矿物表面的吸附量与金属浸出率的关系,必须测定矿物表面吸附的细菌数量.在100℃的水浴中,用0.5mol/L的NaOH溶液消化矿物表面的细菌时间为25min,然后用0.5mol/L的HCl溶液中和至pH=7.0.在2mL中和液中加入1mL茚三酮显色液,在100℃的水浴中加热20min,冷却6min后,在波长为562nm时测定反应产物的吸光值A.吸光值A对应溶液中的蛋白质含量,进而对应着矿物表面吸附细菌的数量.     

镉(Ⅱ)- 蛋白质衍生物的极谱吸附波与应用

研究了镉 ( ) -蛋白质衍生物在氨基乙酸 -氢氧化钠缓冲介质中的极谱行为 ,提出了一种灵敏的蛋白质测定的新方法 .含有巯基或二硫键蛋白质在 0 .2 m ol/ L 氢氧化钠溶液中于沸水浴加热 5 min后 ,在 0 .4 m ol/ L 氨基乙酸 -氢氧化钠 (p H=9.8)缓冲介质中 ,与 1 .0× 1 0 - 4m ol/ L 镉 ( )溶液混合 ,反应产物在单扫描极谱上于 - 0 .5 7V(vs.SCE)处产生一个稳定的、灵敏的吸附还原波 .峰电流与牛血清白蛋白 (BSA)、人血清白蛋白 (HSA)的浓度在7.5× 1 0 - 1 0 ～ 3.0× 1 0 - 7m ol/ L范围内呈线性关系 ,BSA、HSA检测限均为 3.0× 1 0 - 1 0 m ol/ L ;与溶菌酶 (L yso)的浓度在 3.5× 1 0 - 9～ 1 .4× 1 0 - 6 m ol/ L范围内呈线性关系 ,L yso的检测限为 1 .4× 1 0 - 9m ol/ L .该法应用于人血清样品中蛋白质的直接测定 ,结果满意     

利用农业废弃物微生物共发酵制取蛋白质的研究

采用宇佐美曲霉等4种真菌与热带假丝等酵母菌对农业废弃物秸秆进行混合发酵制取蛋白,产物中必需氨基酸含量高于联合国粮农组织(FAO)规定标准,蛋白含量超过玉米和小麦,可作为动物的蛋白饲料.     

Protein tyrosine phosphatase is possibly involved in cellular signal transduction and the regulation of ABA accumulation in response to water deficit in Maize L.coleoptile

Water deficit-induced ABA accumulation is an ideal model or “stimulus-response”system to investigate cellular stress signaling in plant cels,using such a model the cellular stress signaling triggered by water deficit was investigated in Maize L.coleoptile.Water deficit-induced ABA accumulation was sensitively blocked by NaVO3,a potent inhibitor both to plasma membrane H^ -ATPase(PM-H^ -ATPase)and protein tyrosine phosphatase(PTPase).However,while PM-H^ -ATPase activity was unaffected under water deficit and PM-H^ -ATPase activator did not induce an ABA accumulation instead of water deficit,water deficit induced an increase in the protein phosphatase activity,and furthermore,ABA accumulation was inhibited by PAO,a specific inhibitor of PTPase.These results indicate that protein phosphtases may be involved in the cellular signaling in response to water deficit.Further studies identifiled at least four species of protein phosphtase as assayed by using pNPP as substrate,among which one component was especially sensitive to NaVO3.The NaVO3-sensitive enzyme was purified and finally showed a protein band about 66kD on SDS/PAGE.The purified enzyme showed a great activity to some specific PTPase substrates at pH 6.0.In addition to NaVO3,the enzyme was also sensitive to some other PTPase inhibitors such as Zn^2 and MO3^3 ,but not to Ca^2 and Mg^2 ,indicating that it might be a protein tyrosine phosphatase.Interestingly,the purified enzyme could be deactivated by some reducing agent DTT.which was previously proved to be an inhibitor of water deficit-induced ABA accumulation.This result further proved that PTPase might be involved in the cellular signaling of ABA accumulation in response to water deficit.     

Protein P37 of mycoplasma hyorhinis induces secretion of TNF-α from human peripheral blood mononuclear cells

High mycoplasmal infection ratio in gastric cancer tissues suggests a possible association between my-coplasma infection and tumorigenesis.Because TNF-α plays an important role in carcinogenesis caused by microbes in-fection and P37 is a major immunogen of mycoplasma hy-orhinis(M.hyor.),investigating whether P37 could induce expression and secretion of TNF-α will be very significant fo elucidate the possible molecular mechanism of gastric car-cinogenesis involved with M.hyor.At the present study,we cloned full gene of p37 by PCR and mutated the 7 codes of TGA into TGG firstly,then expressed the P37 protein suc-cessfully with pGEX-4T-1 vector in E.coli,which was veri-fied with Western bolt.By RT-PCR and sensitive L929 cell toxic assay,we found that P37 protein could induce expres-sion and secretion of TNF-α from human peripheral blood mononuclear cells,and the inducing activity of P37 could be dramatically blocked by McAb PD4.These results suggest that the induction of TNF-α secretion by P37 probably plays an importan role in diseases caused by M.hyor.infection and needs to be further investigated.     

SHRIMP U-Pb zircon geochronology of Yangfang aegiriteaugite syenite in Wuyi Mountains of South China and its tectonic implications

In addition to the wide spread peraluminous granites, some alkaline or alkali-rich intrusive rocks were recently observed in the Wuyi Mountains. The Yangfang aegiriteaugite syenites in the Wuyi Mountains are strongly enriched in light rare earth elements (LREE), K, Ba, Ga, Zr and Y. What is more, differentiations of REE are obvious. In addition, the Yangfang aegiriteaugite syenites have also some characteristics similar to A-type granite. Zircons are selected from the Yangfang aegiriteaugite syenites for Sensitive High-Resolution Ion Microprobe (SHRIMP-Ⅱ) U-Pb geochronology study. 15 analyses give concordant 206Pb/238U ages in a narrow range of 233—249 Ma, which correspond to a single age population with a weighted mean 206Pb/238U age of 242 4 Ma, which records the crystallization time of the aegiriteaugite syenites. It is suggested that: (1) there were early Triassic peralkaline intrusive rocks in South China; (2) the early Indosinian extension event was probably active in local area, South China. This discovery provides a new insight for farther understanding of 揑ndosinian orogeny in South China.     

HMM in predicting protein secondary structure

We introduced a new method—duration Hidden Markov Model (dHMM) to predicate the secondary structure of Protein. In our study, we divide the basic second structure of protein into three parts: H (α-Helix), E (β-sheet) and O (others, include coil and turn). HMM is a kind of probabilistic model which more thinking of the interaction between adjacent amino acids (these interaction were represented by transmit probability), and we use genetic algorithm to determine the model parameters. After improving on the model and fixed on the parameters of the model, we write a program HMMPS. Our example shows that HMM is a nice method for protein secondary structure prediction. Foundation item: Supported by the National Natural Science Foundation of China (30170214) Biography: Huang Jing (1977-), female, Master candidate, research direction: bioinformatics.     

Inhibitory effect of the adenovirus type 5 E1A protein expressed in the yeast system on the human tumor cell growth

Adenovirus 5 type E1A as a tumor suppressor gene can inhibit tumor growth and enhance the sensitivity of chemotherapy and radiotherapy. E1A have the ability to integrate into the host genome, resulting in long-time expression that induces Rb gene inactivation and animal cells immortalization. This prompted us to select the E1A protein for treatment of cancer in order to overcome the limitations of E1A gene therapy. Thus, we firstly constructed E1A eucaryotic expression vector (pPIC9/E1A), transformated the pichia pastoris yeast cells (GS115) and screened the high-expressing recombinant strains. The positive yeast strains were cultured in the shake flask, and induced for 3 d. The crude E1A protein was purified using two steps of column chromatography on HiTrap Q and HiTrap SP. The purified E1A protein was identified by SDS-PAGE and Western blot. E1A protein was mostly located at cellular nuclear when Chariot delivered E1A protein into cells. The analysis in vitro indicated that the E1A protein arrested LN686 cell cycle at G2/M phase, and significantly inhibited the growth of LN686 tumor cells. The current studies firstly provided an experimental basis to further develop E1A protein for tumor treatment.     

Identification and characterization of a salt tolerance-responsive gene( AtGRP9) of Arabidopsis

Soil salinity is one of the important limiting factors for plant growth and development. A cDNA clone encoding a glycine-rich protein (designated AtGRP9) was identified from Arabidopsis by functional expression of the plant cDNA library in the fission yeast S. pombe. Yeast cells overexpressing AtGRP9 displayed significantly enhanced salt tolerance. Northern analysis showed that expression of AtGRP9 in Arabidopsis was induced by NaCl and plant hormone abscisic acid (ABA). These results suggest that AtGRP9 may be involved in the salt stress response in Arabidopsis.     

Heterotrimeric G protein participated in modulation of cytoplasmic calcium concentration in pollen cells

Cytoplasmic free calcium concentration([Ca2+]c) in pollen cells of Lilium daviddi is measured with confocal laser scanning microscopy to investigate the effect of heterotrimeric G protein (G protein) on [Ca2+]c and the possible signal transduction pathway of G protein triggering cellular calcium signal. After application, cholera toxin (CTX), an agonist of G protein, triggers a transient increase of [Ca2+]c in pollen cells, and evokes a spatial-temporal characteristic calcium dynamics; while pertussis toxin (PTX), a G protein antagonist, leads to the decrease of [Ca2+]c. Both L-type Ca2+ channel blocker verapamil and inhibitor of IP3 receptor heparin inhibit CTX-induced [Ca2+]c increase. The results show that G protein may play a role in the modulation of [Ca2+]c through enhancing the extracellular Ca2+ influx and releasing of Ca2+ from intracellular stores.