Direct electrochemical identification of an activated intermediate formed by cytochrome C with hydrogen peroxide

An activated intermediate formed from H2O2 and cytochrome C is identified by direct electrochemical measurements.

Expression of beta-carotene hydroxylase gene (crtR-B) from the green alga Haematococcus pluvialis in chloroplasts of Chlamydomonas reinhardtii

A carotenoid gene (crtR-B) from the green alga Haematococcus pluvialis, encoding beta-carotene hydroxylase that was able to catalyze the conversion of beta-carotene to zeaxanthin and canthaxanthin to astaxanthin, was cloned into Chlamydomonas reinhardtii chloroplast expression vector p64D to yield plasmid p64DcrtR-B. The vector p64DcrtR-B was transferred to the chloroplast genome of C. reinhardtii using micro-particle bombardment. PCR and Southern blot analyses indicated that crtR-B was integrated into the chloroplast genome of the transformants. RTPCR assays showed that the H. pluvialis crt R-B gene was expressed in C. reinhardtii transformants. The transformants rapidly synthesized carotenoids in larger quantities than the wild-type upon being transferred from moderate to high-intensity white light. This research provides a foundation for further study to elucidate the possible mechanism of photo-protection by xanthophylls and other carotenoids in high light conditions or through exposure to UV radiation.

小麦春化相关基因的克隆及其功能分析

一． 小麦相关基因ver203F cDNA全序列克隆与功能分析 　　根据本实验室通过差异筛选技术克隆到的与春化相关的基因cDNA verc203的序列，设计PCR 5’端PCR引物，利用RACE（rapid amplification of cDNA ends）克隆策略，得到春化相关基因ver203基因的同源基因ver203F cDNA的3’端序列，长度为1,197bp。Northern分析表明ver203F全长约为1.5 kb，且其表达具有春化处理的特异性。根据3’RACE克隆的ver203F 3 ’端核苷酸序列设计了3’端PCR引物，利用5’RACE克隆到该基因的5’端片段，经DNASIS核酸分析软件分析将5’45RACE和3’RACE DNA序列拼接合并，得到ver203F全长cDNA，从TdT加尾5’末端到poly A全长为1,561 bp，5’端起始密码子ATG上游非编码区－1～－192共了192bp，终止密码子TGA到poly A的非编码区有253bp，cDNA编码区全长1,119 bp，推测编码373氨基酸残基。国际基因序列数据库检索表明该基因序列（GenBank/EMBL/DDBJ：AB012013）与大麦茉莉酸诱导基因有部分同源性。因上推测该基因在调控开花过程中可能参与茉莉酸介导的信号传导途径，ver203F作用的发挥可能需要其它蛋白的参与，或ver203F本身就是一个受体蛋白。 　　为了研究ver203F基因的功能，将通过3’RACE克隆到的ver203F 3’端序到分别构建正义和反义植物表达载体，通过花粉管通道法、农杆菌介导的叶圆片法以及农杆菌介导的真空转化法分别转化小麦、烟草和拟南芥菜。获得转基因植株后，PCR、DNA Dob Blot、Southern Blot分析以及GUS活性检测证明外源基因已整合到转基因植株中，并得到表达。在获得的小麦、烟草和拟南芥菜转基因植株中，它们开花时间都相应地推迟，表明正常植物体内该基因在控制营养生长向生殖生长的转变中起作用。ver203F可以影响小麦和拟南茶菜花序的发育，首先无论正义还是反义都使得花序的发育受到抑制，在小麦中表现为顶部小穗退化，在拟南芥菜中表现为顶花。其次在转化正义基因的转基因拟南芥菜中，观察到产生的顶花为对称的两朵或以对称的两朵顶花基部为生长点长出丛生花，这种对称花的麦型小麦小穗中小花的表型相类似，说明ver203F基因可能在小麦小花的发育过程中也起着重要作用。 二． 春化相关基因ver17在开花过程中功能的分析 以春化处理冬小麦（京冬1号）幼苗cDNA为材料，通过减法杂交与差异筛选得到春化相关cDNA克隆verc17。为了研究该基因的功能，以包含CaMV 35S启动子的pBI221为载体，将ver17cDNA片段分别从两个方向插入pBI221的BamH I－Sma I, Xba I－BamH I间，构建正义和反义表达质粒：p17S和p17X，通过花粉管通道法转化小麦。对T0和T1两代转基因小麦的观察发现，在转化反义基因p17X的转基因小麦首先表现为抽穗推迟，其次穗的顶部和基部小穗严重退化，另外还发现转化反义基因的小麦败育现象严重（主要是花粉败育），因此推测ver17基困能可具有以下几方面的特点：a．春化诱导型表达;b．促进植物开花;c．促进穗顶端和基部花的发育，减少其退化;d．影响雄蕊的发育。

通过第一性原理计算研究内地核的结构与成分

The determination of the composition and structure of the Earth’s inner core has long been the major subject in the study of the Earth’s deep interior. It’s widely believed that the Earth’s core is formed by iron with a fraction of nickel. However, light elements must exist in the inner core because the earth core is less dense than pure iron-nickel alloy (~2-3% in the solid inner core and ~6-7% in the liquid outer core). The questions are what and how much light element is there in the iron-nickel alloy. Besides the composition, the crystal structure of the iron with or without light element is also not well known. According to the seismological observations, the sound waves propagate 3-4% faster along the spin axis than in the equatorial plane. That means the inner core is anisotropic. The densest structure of iron-nickel alloy should be h.c.p structure under the very high pressures. However, the h,c,p structure does not propagate waves anisotropic ally. Then what is the structure of the iron-nickel alloy or the iron-nickle-light element alloy. In this study, we tried to predict the composition and the structure of the inner core through ab initio calculation of the Gibbs free energy, which is a function of internal energy, density and entropy. We conclude that the h.c.p structure is more stable than the b.c.c structure under high pressure and 0 K, but with the increase of temperature, the free energy of the b.c.c structure is decreasing much faster than the h.c.p structure caused by the vibration of the atomics, so the b.c.c structure is more stable at high temperatures. With the addition of light elements (S or Si or both), the free energy of b.c.c. decreases even faster, about 3at% of Si not only explains why the inner core is about 2-3 % lighter than the iron-nickle alloy, but also reasons why the inner core is anisotropic, since the b.c.c. structure becomes more stable than the h.c.p structure at 5500-6000K and b.c.c. is anisotropic in propagating seismic waves. Therefore, we infer that the inner core of the earth is formed by b.c.c iron and a fraction of nickel plus ~3at.% Si, with a temperature higher than 5500K, which is consistent with the studies from other approaches.

济阳-昌潍坳陷火成岩油气藏体系形成机理和分布规律研究

Jiyang & Changwei depressions are two neighboring depressions in Bahai Bay Basin, the famous oil rich basin in East China. The exploration activities in the past 40 years has proved that, within the basins, there exists not only plentiful sandstone hydrocarbon reservoirs (conventional), but also abundant special reservoirs as igneous rock, mudstone and conglomerate ones which have been knowing as the unconventional in the past, and with the prospecting activity is getting more and more detailed, the unconventional reservoirs are also getting more and more important for further resources, among which, the igneous lithological reservoir be of significance as a new research and exploration area. The purpose of this paper is, with the historical researches and data as base, the System Theory, Practice Theory and Modern Comprehensive Petroleum Geology Theory as guide, the theoretical and practice break through as the goal, and the existing problems in the past as the break through direction, to explore and establish a valid reservoir formation and distribution models for igneous strata in the profile of the eastern faulted basins. After investigating the distribution of the igneous rocks and review the history of the igneous rocks reservoirs in basins, the author focused on the following issues and correspondingly the following progresses have been made: 1.Come to a new basin evolution and structure model named "Combined-Basin-bodies Model" for Jiyang even Eastern faulted basins based on the study on the origin and evolution of Jiyang & Changwei basins, depending on this model, every faulted basin in the Bo-hai Bay Basin is consisted of three Basin-Bodies including the Lower (Mesozoic), Middle (Early Tertiary) and the Upper (Late Tertiary) Bodies, each evolved in different geo-stress setting and with different basin trend, shape and igneous-sedimentary buildings system, and from this one to next one, the basin experienced a kind of process named "shape changing" and "Style changing". 2. Supposed a serious of new realizations as follows (1) There were "multi-level magma sources" including Upper mantel and the Lower, Middle and even the Upper Shell magma Chambers in the historical Magma Processes in the basins; (2) There were "multi-magma accessing or pass" from the first level (Mantel faults) to the second, third and fourth levels (that is the different levels of fault in the basin sediment strata) worked in the geo-historical and magma processes; (3) Three tectonic magma cycles and more periods have been recognized those are matched with the "Basin -body-Model" and (4)The geo-historical magma processes were non-homogeneous in time and space scale and so the magma rocks distributed in "zones" or "belts". 3. The study of magma process's effect on basin petroleum conditions have been made and the following new conclusions were reached: (1) the eruptive rocks were tend to be matched with the "caped source rock", and the magma process were favorable to the maturing of the source rocks. (2) The magma process were fruitful to the accumulation of the non-hydrocarbon reservoirs however a over magma process may damage the grade of resource rock; (3) Eruptive activity provided a fruitful environment for the formation of such new reservoir rocks as "co-eruptive turbidity sandstones" and "thermal water carbonate rocks" and the intrusive process can lead to the origin of "metamorphism rock reservoir"; (4) even if the intrusive process may cause the cap rock broken, the late Tertiary intrusive rocks may indeed provide the lateral seal and act as the cap rock locally even regionally. All above progresses are valuable for reconstructing the magma-sedimentary process history and enriching the theory system of modem petroleum geology. 4. A systematic classification system has been provided and the dominating factors for the origin and distribution of igneous rock reservoirs have been worked out based on the systematic case studies, which are as follows: (1) The classification is given based on multi-factors as the origin type, litho-phase, type of reservoir pore, reservoir ability etc., (2) Each type of reservoir was characterized in a detailed way; (3) There are 7 factors dominated the intrusive reservoir's characteristics including depth of intrusion, litho-facies of surrounding rocks, thickness of intrusive rock, intrusive facies, frequency and size of the working faults, shape and tectonic deformation of rock, erosion strength of the rock and the time of the intrusion ect., in the contrast, 4 factors are for eruptive rocks as volcanic facies, frequency and size of the working faults, strength of erosion and the thermal water processing. 5. Several new concept including "reservoir litho-facies", "composite-volcanic facies" and "reservoir system" ect. Were suggested, based on which the following models were established: (1) A seven reservoir belts model for a intrusive unit profile and further more, (2) a three layers cubic model consisted of three layer as "metamorphic roe layer", "marginal layer" and "the core"; (3) A five zones vertical reservoir sequence model consisted of five litho-facies named A, B, C, D and E for a original lava unit and furthermore three models respectively for a erosion, subsidence and faulted lava unit; (4) A composite volcanic face model for a lava cone or a composite cone that is consisted of three facies as "crater and nearby face", "middle slope" and "far slope", among which, the middle slope face is the most potential reservoir area and producible for oil & gas. 6. The concept of "igneous reservoir" was redefined as the igneous, and then a new concept of "igneous reservoir system" was supposed which means the reservoir system consisted of igneous and associated non-igneous reservoirs, with non-hydrocarbon reservoir included. 7. The origin and distribution of igneous reservoir system were probed and generalized for the exploration applications, and origin models of the main reservoir sub-systems have been established including those of igneous, related non-igneous and non-hydrocarbon. For intrusive rocks, two reservoir formation models have been suggested, one is called "Original or Primary Model", and the another one is "Secondary Model"; Similarly, the eruptive rock reservoirs were divided in three types including "Highly Produced", "Moderately Produced" and "Lowly Produced" and accordingly their formation models were given off; the related non-igneous reservoir system was considered combination of eight reservoirs, among which some ones like the Above Anticline Trap are highly produced; Also, the non-hydrocarbon. Trap system including five kinds of traps was discussed. 8. The concept models for four reservoir systems were suggested, which include the intrusive system consisted of 7 kinds of traps, the land eruptive system with 6 traps, the under water eruptive system including 6 kinds of traps and the non-hydrocarbon system combined by 5 kinds of traps. In this part, the techniques for exploration of igneous reservoir system were also generalized and probed, and based on which and the geological progresses of this paper, the potential resources and distributions of every reservoir system was evaluated and about 186 millions of reserves and eight most potential non-hydrocarbon areas were predicted and outlined. The author believe that the igneous reservoir system is a very important exploration area and its study is only in its early stage, the framework of this paper should be filled with more detailed studies, and only along way, the exploration of igneous reservoir system can go into it's really effective stage.