Three 5’-flanking Regions of crtO Encoding β-carotene Oxygenase in Haematococcus pluvialis

Three separate 5＇-flanking regions （1.1 kb, 1.9 kb and 2.2 kb） of crtO were cloned through walking upstream. Results of sequence analysis show that three 5＇-flanking regions of crtO might have some similar putative cis-acting elements such as ABA （abscisic acid）-responsive element （ABRE）, C-repeat/dehydration responsive element （C-repeat/DRE）, light-responsive element （G-box, GAG-motif, I-box and ATC-motif）, wound-responsive element （WUN-motif）, auxin-responsive element （TGA-element）, MeJA-responsive element （TGACG-element） and MYB binding site （MBS）, except for typical TATA box or CCAAT box. These findings might mean diversiform regulatory patterns of crtO being in astaxanthin biosynthesis of Haematococcus pluvialis.

Sequence variability of the 5, UTR in isolates of hepatitis C virus in China

Background: Hepatitis C virus infection is a great is- sue in China; however, there is very little informa- tion on genotyping investigations based on sequence variability in the 5' untranslated (5'UTR) reported. The present study was to define the sequence varia- bility based on the sequence divergences of the 5' UTR of the virus. Methods: Sequences of 91 isolates from patients with chronic hepatitis C from Yunnan, southwest China, were sequenced and genotypes were defined accord- ing to the sequence divergences of the 5' UTR of the virus. Results: Eighty-six isolates were classified into 3 clades (previously termed groups or major types) by the methods proposed by Chan et al in 1992 and phy- logenetic analysis based on nucleotide sequence diver- gences within the 5' UTR. Fifty-six percent of the i- solates were classified into clade 3, 35% into clade 1, and 34.9% into clade 2. New genotypes 1f, 2h, 3h and 3i were defined. In addition, 3 novel sequences were discovered, respectively with an 18-nt sequence deletion (corresponding to nucleotide position -173 to -156), a 28-nt sequence insertion, and a 40-nt se- quence insertion, between -56 and -55. Of these i- solates, 56% possessed a 'G' at position -66 in place of the 'T' that is present in all previously re- ported sequences. Conclusions: These HCV variants, evolved or re- mained in this area, may be of great significance in diagnosis and treatment of hepatitis C patients.

Grain-size Characteristics and Climate Variability in TMS5e Sequence of Tumen Section in Southern Tengger Desert,Northwestern China

The TMS5e sequence from the Tumen section, at the southern edge of Tengger Desert in the northwestern China, is synchronous with Marine Isotope Stage 5e （MIS5e）. It consists of 16 layers of aeolian dune sands, 11 layers of lacustrine loess-like facies, and 5 layers of lacustrine facies. The results of grain-size analysis shows that the pa- laeo-mobile dune sands, palaeo-fixed to semi-fixed dune sands and loess-like sandy loams are mainly composed of sands, ranging from 70% to 96%; their silt contents ranged from 4% to 20%, and their clay contents ranged from 1% to 5%; the climate under which the aeolian dune sands were deposited is similar to that under which modern mobile dune sands form, which is caused by the dominance of the cold, dry East Asian winter monsoon. In contrast, the lacustrine loess-like facies and lacustrine facies had a lower sand contents than those of the three aeolian dune sands, but have higher silt and clay contents, most of their sand content ranged from 30% to 60%, their silt contents ranged from 35% to 55%, and their clay contents ranged from 6% to 20%. The lacustrine loess-like facies and lacustrine facies formed under the influence of the warm, humid East Asian summer monsoon based on their similarity with modern sediments. The grain-size indicator Mz （mean grain diameter） and the SC/D value in the TMS5e sequence indicate climatic insta- bility at the southern edge of the Tengger Desert during MIS5e, with at least 14 fluctuations between a warm, humid climate and a cold, dry climate, divided into five stages： TMS5e5 （139 kyr to 129.3 kyr B.P.）, TMS5e4 （129.3 kyr to 124 kyr B.P.）, TMS5e3 （124 kyr to 119.5 kyr B.P.）, TMS5e2 （119.5 kyr to 116.5 kyr B.P.）, and TMS5el （116.5 kyr to 113.7 kyr B.P.）. These correspond roughly to MIS5e5, MIS5e4, MIS5e3, MIS5e2, and MIS5el, respectively, in the GRIP ice core data.

Effect of Impregnation Sequence on Propane Dehydrogenation Performance of PtSnNa/ZSM-5 Catalyst

The effects of the sequence for impregnation of metal precursors on the performance of PtSnNa/ZSM-5 catalyst for propane dehydrogenation to propene were studied in this paper.Some methods such as XRD,TPDA,BET,H2-TPR,XPS,ICP,TEM and hydrogen chemisorption were used to characterize the catalysts.The structure of ZSM-5 zeolite was not destroyed by the introduction of metal components.Meanwhile the different impregnation sequence of metal precursors could affect the behavior of Sn4+species entering the ZSM-5 channel,and the interaction between platinum and tin species,as well as the degree for reduction of Pt and Sn components.As a result,the prepared catalysts exhibited different reaction activity and selectivity.Compared with the co-impregnation treated catalyst,the catalysts prepared by the sequential impregnation method showed better catalytic activity in propane dehydrogenation,especially the one prepared through impregnation with tin precursor at first.Finally,a model for the effect of impregnation sequence on the distribution of Pt and Sn species in PtSnNa/ZSM-5 catalyst was proposed.

THE ABNORMAL EXPRESSION OF CLONED REPEATED SEQUENCE DNA, L5B-4, IN RAT HEPATOMA BERH-2

A repeated sequence DNA fragment, L5B-4, was cloned from the 5 kb BamHI DNA fragments of rat genomic DNA. The expressions of the L5B-4 DNA fragment are different in liver and hepatoma cells. The amounts of transcripts in hepatoma cells are lower in nucleus and higher in cytoplasm, especially in polysomal RNA, as compared with that in liver cells. The alteration shown in polysomal RNA of hepatoma cells seems to be specific. These results are discussed with respect to the possible function of this repeated DNA and its variation in hepatoma cells.

The Reaction Sequence and Dielectric Properties of BaAl₂Ti₅O₁₄Ceramics

To investigate the correct reaction sequence of BaO-Al2O3-5TiO2 system, powders calcined at different temperatures are analyzed by x-ray diffraction. The results show that the source phase BaCO3 decomposes below 800°C, TiO2 and Al2O3 start to consume at 900 and 1100°C, respectively. BaTi4O9 phase appears at 1000°C while BaAl2Ti5O14 phase starts to reveal at 1200°C. As the temperature increases, the density, dielectric constant and quality factor of the BaAl2Ti5O14 ceramic increase and keep unchanged at 1350°C. The dielectric properties of BaAl2Ti5O14 ceramic sintered at 1350°C for 3h are: εr=35.8, Q×f=5130GHz, τf=-6.8ppm/°C.

Cloning and Sequence Analysis of rbcS Gene of Wild Barley (Hordeum brevisubulatum) under Salt Stress

[Objective] The aim was to study the cloning and sequence analysis of rbcS gene of wild barley under salt stress. [Method] The tender leaf blade of wild barley under salt stress was taken as the experimental material. The primers were designed according to the homology of rbcS gene sequences of wheat and barely in Genbank; then PCR amplification,recovery,ligation,transformation and sequencing of rbcS gene were carried out. [Result] Two rbcS genes including rbcS1 and rbcS2 with the length of 1 252 and 908 bp respectively were cloned from the barely genome. rbcS1 and rbcS2 were both composed by two exons and one intron. The exons length of the two genes was the same of 525 bp,encoding 174 amino acids,and the homology between them was 96%; however,the intron length of rbcS1 and rbcS2 was 448 and 107 bp respectively.

Small cell lung carcinoma with KIF5B-RET fusion partially responded to the 4^(th)-line therapy with anlotinib:A case report

BACKGROUND Small cell lung cancer(SCLC)exhibits a pronounced tendency for metastasis and relapse,and the acquisition of resistance to chemotherapy and radiotherapy,leading to complexity in treatment outcomes.It is crucial to tackle these challenges by advancing targeted therapeutic approaches in ongoing research endeavors.Variant RET fusions have been reported in several solid tumors,but are rarely reported in SCLC.CASE SUMMARY We present the first case of a KIF5B-RET fusion in a 65-year-old male patient with SCLC.To date,the patient has received the 4th line chemotherapy with anlotinib for one year and has shown a sustained favorable partial response.According to the results of next generation sequencing,this SCLC patient harbors the KIF5BRET fusion,suggesting that RET fusion could serve as a promising molecular target for SCLC treatment.Next-generation sequencing(NGS)plays a critical rolein comprehensively assessing the genotype and phenotype of cancer.CONCLUSION NGS can provide SCLC patients with personalized and targeted therapy options,thereby improving their likelihood of survival.

血清FAM19A5、IL-34在血管性认知障碍患者中的表达及临床意义

目的探讨血清序列相似家族19成员5(FAM19A5)、白介素-34(IL-34)在血管性认知障碍(VCI)患者中的表达及临床意义。方法选取2020年1月—2022年12月包头市中心医院神经内科收治的非痴呆血管性认知损害(VCIND)患者125例(VCIND组)和同期体检健康者58例(健康对照组),VCIND组患者随访1年根据是否进展为血管性痴呆(VaD)分为VaD亚组45例和非VaD亚组80例,采用酶联免疫吸附法检测血清FAM19A5、IL-34水平。通过多因素Logistic回归分析VCIND进展至VaD的影响因素,ROC曲线分析血清FAM19A5、IL-34水平对VCIND进展至VaD的预测能效。结果与健康对照组比较,VCIND组血清FAM19A5水平升高,IL-34水平降低(t=19.808、15.706,P均<0.001)。随访1年,125例VCIND患者VaD发生率为36.00%(45/125)。与非VaD亚组比较,VaD亚组血清FAM19A5水平升高,IL-34水平降低(t=7.100、5.952,P均<0.001)。Logistic回归分析结果表明,年龄增加(OR=1.137,95%CI 1.014~1.275)、FAM19A5升高(OR=1.079,95%CI 1.040~1.119)为VCIND进展至VaD的独立危险因素(P<0.05),蒙特利尔认知评估量表评分增加(OR=0.603,95%CI 0.437~0.832)、IL-34升高(OR=0.737,95%CI 0.624~0.870)为独立保护因素(P<0.05)。血清FAM19A5、IL-34水平联合预测VCIND进展至VaD的曲线下面积为0.880,大于血清FAM19A5、IL-34水平单独预测的0.813、0.781(Z=2.529、2.855,P均<0.05)。结论血清FAM19A5水平升高和IL-34水平降低与VCIND进展至VaD密切相关,血清FAM19A5、IL-34水平联合检测对VCIND进展至VaD有较高的预测效能。

Analysis on Heredity and Variation of the ORF_5 Gene of Prevalence Strains Porcine Reproductive and Respiratory Syndrome Virus

[ Objective] The aim was to analyze the reason and epidemic trend of PRRSV, and provide theoretical basis for preventing and controlling PRRS. [Methed]According to the sequence of ATCC VR-2332 strain porcine reproductive and respiratory syndrome virus published by the GenBank, the primers were designed and synthesized. ORF5 gene sequences of seven prevalence strains were amplified by RT-PCR. The sequences of ORF5 genes were analyzed by DNAStar and compared with those of ATCC VR-2332, CH-1 a, B J-4, LV-M96262 and MLV vaccine strains, phylogenetic tree among isolates was analyzed. [Result] Analysis of nucleotide sequence showed that the homology was 88.1% - 98.8%, 89.9% -95.2%, 85.6% -98.7% between ORF5 genes of seven prevalence strains and VR-2332, CH-1a, BJ-4, and the homology was 54.7% -56.9% between ORF5 genes and LV. Analysis of amino acid sequence showed that the homology was 88.1% -96.8%, 88.1% - 94.5%, 86.1% -96.5% between ORF5 genes of seven prevalence strains and VR-2332, CH-1a, bBJ-4, the homology was 54.7% -56.2% between the ORF5 genes and LV.[ Conclusion] The variation of prevalence strains was great in the ORF5 gene region, the homology of ORF5 gene sequence was higher and genetic relationship was nearer during prevalence strains in the same region, or was far in different regions.

牛病毒性腹泻病毒长春分离株P125基因的克隆与序列测定

以MDBK细胞培养致细胞病变型牛病毒性腹泻病毒 (BVDV)NCD毒株 ,采用异硫氰酸胍一步法提取总RNA ,并根据BVDVNADL参考株序列设计合成的 1对引物 (W 1与W 2 ) ,进行反转录PCR (RT PCR) ,扩增出P12 5基因区约40 0bp的片段。经 pGEM T载体连接后克隆、测序 ,并与已发表的NADL、Osloss、SD 1、184、D、H、Yak等BVDV毒株序列进行比较。结果表明 :NCD株P12 5基因区无插入或缺失变异 ,但存在着核苷酸的替换 ;经聚类分析初步认为NCD株属于Ⅰa型。NCD株与长春 184、H株核苷酸序列差异较大 ,而亲缘关系较近 。

鳜鱼(Siniperca chuatsi)β-肌动蛋白基因cDNA全序列与5′侧翼区的克隆与分析

采用RT-PCR及RACE法,分离、克隆鳜鱼β-肌动蛋白基因cDNA全序列,再用基因组步行法(Genome Walker)克隆鳜鱼β-肌动蛋白基因5′调控区。序列分析结果表明,鳜鱼β-肌动蛋白基因全长1897bp,其中5′-UTR长94bp,3′-UTR长675bp,编码区长1128bp,编码375个氨基酸。将所得序列与其它动物类群的β-肌动蛋白基因序列进行比较分析显示,鱼类、两栖类、鸟类、哺乳类等不同类群脊椎动物β-肌动蛋白氨基酸序列同源性均在96%以上,说明该基因在生物进化过程中高度保守。通过鳜鱼与其它脊椎动物β-肌动蛋白基因的核苷酸序列构建的进化树显示,脊椎动物β-肌动蛋白聚类成3个分支,鱼类β-肌动蛋白基因形成一个独立的分化群,说明鱼类β-肌动蛋白基因起源于一个共同祖先。克隆得到的鳜鱼β-肌动蛋白基因5'侧翼序列长1399bp,对其进行序列分析,在其起始密码字ATG上游200bp范围内发现含有CAATbox、CC(A/T)6GG(CArGbox)、TATA box对转录调控起重要作用的顺式元件,同时在侧翼区也发现含有GC box、MYOD、YY1、SP1、GATA等多个潜在调控元件。鳜鱼β-肌动蛋白基因5′侧翼序列的克隆成功,为今后转基因鳜鱼的研究工作奠定了基础。

利用5′锚定PCR技术分离枇杷微卫星标记

用5′锚定PCR技术分离出32个枇杷特异SSR位点,对其中适合设计引物的28个位点设计了相应的引物,并分别与对应的5′锚定的简并SSR引物配对,应用于5个枇杷品种的基因组PCR扩增。结果表明,27条引物在5个受试枇杷品种中有扩增产物,其中24条为可用引物,另外3条引物因为扩增产物不符合SSR标记的特征不能用作SSR标记的引物。

柠条GDP-甘露糖-3′,5′-异构酶基因的克隆、序列分析及其植物表达载体的构建

L-抗坏血酸是一种抗氧化剂,能够保护植物免受活性氧的伤害,GDP-甘露糖-3′,5′-异构酶基因(GME)是L-抗坏血酸合成途径中的关键基因。根据其他植物GME的氨基酸保守序列设计简并引物,通过RT-PCR结合RACE扩增技术,从柠条叶片中克隆了1个编码GME的基因,命名为CkGME,Genebank登录号为FJ603689。基因全长1604bp,开放阅读框1134bp,编码377个氨基酸,氨基酸同源性比对分析表明,该基因编码的氨基酸与其他植物GME基因具有较高的同源性。进化树分析表明,该基因编码的氨基酸序列与豆科植物大豆和截型苜蓿的亲缘关系最近,与十字花科植物拟南芥的亲缘关系次之,与禾本科植物水稻和玉米的亲缘关系较远。利用4℃和GA3处理不同时间的柠条植株进行半定量RT-PCR分析结果表明,该基因在4℃处理不同时间后表达量没有明显变化,在GA3处理不同时间后表达量显著下降。利用pCAMBIA-1300质粒构建含35S启动子的正义表达载体,酶切图谱和PCR检测结果证明该表达载体构建成功。

b-Boule基因5′调控序列的克隆与睾丸组织DMR甲基化分析

【目的】研究b-Boule基因5′调控序列的序列特征,以及牦牛、黄牛与犏牛睾丸组织b-Boule基因DMR甲基化状态的差异,为揭示b-Boule基因的表达调控和犏牛雄性不育的表观遗传机制提供依据。【方法】采用PCR扩增和克隆测序技术获得牦牛b-Boule基因5′调控序列,利用生物信息学方法分析b-Boule基因5′调控序列的序列特征,采用亚硫酸氢钠测序法检测牦牛、黄牛与犏牛睾丸组织中b-Boule基因DMR的甲基化状态。【结果】b-Boule基因5′调控序列长度为1 352 bp,核心启动子区含有SP1等甲基化敏感位点,5′端存在一个CpG岛。犏牛b-Boule基因DMR的甲基化水平(17.78%)高于牦牛(7.50%)和黄牛(6.94%)(P<0.01),特别是CpG位点33—35的甲基化水平差异更明显。【结论】犏牛b-Boule基因DMR的甲基化水平高于牦牛和黄牛,结合前期mRNA表达水平和组织学观察结果,认为DMR甲基化在b-Boule基因的表达调控中发挥关键作用,犏牛b-Boule基因可能是通过DMR区的高甲基化抑制其mRNA表达来阻滞精子发生减数分裂过程。

果蝇程序化死亡基因5(PDCD5)同源cDNA的克隆和序列分析

为了解人类白血病细胞凋亡相关新基因 TFAR1 9(PDCD5,programmed cell death5)在不同种属间的序列同源性 ,利用 EST(expression sequence tag)拼接、RT- PCR、DNA序列测定技术及计算机分析技术 ,首次成功地进行了果蝇 PDCD5同源 c DNA编码区基因克隆和序列分析 .发现果蝇与小鼠及果蝇与人 PDCD5在核苷酸水平上分别有 57.5%和 57.1 %的同源性 ,在氨基酸水平上分别有 46.8%和 46.4%的同源性 .功能区分析发现 ,果蝇 PDCD5c DNA编码 1 33个氨基酸 ,计算机预测可能是一种核蛋白 ,含 5个可能的酪蛋白激酶 (casein kinase )磷酸化位点 ,2个可能的 PKC磷酸化位点 ,与人 PDCD5的功能区类似 .因而果蝇 PDCD5是与人 PDCD5同源的新基因 ,可能都与细胞程序化死亡相关 .

我国两株登革2型病毒5′和3′端非编码区序列测定及二级结构分析

为测定我国两株临床症状、乳鼠神经毒力不同的登革 2型病毒流行株 5′和 3′端非编码区序列 (untranslated region,UTR) ,分析二级结构差异与毒力变化的关系 ,分别从 D2 - 0 4、D2 - 44株感染的 C6/ 36细胞及鼠脑中提取总 RNA.以该 RNA为模板 ,利用 RACE法 ,分别扩增了 D2 - 0 4、D2 -44株的 5′和 3′末端 c DNA片段 .将其分别与 p GEM- T载体连接得到重组质粒 ,测定上述 c DNA插入片段的序列 .用 RNAdraw软件预测 D2 - 0 4、D2 - 44株 5′和 3′端非编码区的二级结构 .D2 - 0 4、D2 -44株 5′端和 3′端非编码区分别有 96和 454个核苷酸 .其中 5′非编码区 59位 C(D2 - 0 4 )→T(D2 -44 ) ,使 D2 - 44二级结构稳定性下降 ;3′端非编码区有 1 5个核苷酸不同 ,其中 T(355)→ A,T(32 6)→ G引起了所在位置二级结构自由能变化 ,且分别位于两个保守序列区 (conserved sequence,CS)CS1、CS2 A.这些位点变化可能与毒力有关 .

鹅催乳素受体基因cDNA5′端序列克隆

为寻找鹅就巢性相关基因,从洪泽湖性成熟公鹅睾丸组织分离并合成鹅cDNA第一链;利用RACE克隆了鹅催乳素受体基因(gPRLR)5′端序列。所克隆的499 bp 5′端序列可划分为348 bp的5′-UTR、151 bp的以ATG作为起始密码子的阅读开放框。与鸡催乳素受体基因(cPRLR)cDNA序列作比对,此499 bp序列可划分为238 bp的第1外显子、69 bp的第2外显子、114 bp的第3外显子及81 bp的部分第4外显子,起始密码子位于第3外显子45 bp处。阅读开放框核苷酸序列与cPRLR cDNA同源部位的阅读开放框同源性达到88%,推导的氨基酸序列同源性达到84%。

鄂尔多斯盆地上三叠统延长组长4＋5油层组中的古地震记录

鄂尔多斯盆地上三叠统延长组长4+5油层组湖泊三角洲前缘沉积中同生变形构造十分发育。通过对岩心观察及各种资料的综合分析,认为砂球、砂枕、球—枕构造、微断层递变层、微裂缝、液化卷曲变形等同生变形构造主要是由震积液化作用形成的。震积作用沉积可分为A、B、C、D、E等5个单元,分别组合形成ABCD型、BCD型或CD型、AB型、BCE型或BE型、BEBE型5种沉积序列类型,其中BCD型(CD型)和AB型最为发育。通过对古构造的分析,认为晚三叠世鄂尔多斯盆地拗陷中心的北西向基底"活化"及北西、北东向同生正断层的活动是导致该区古地震形成的主要原因。

鲢鱼可溶性谷胱甘肽S-转移酶(sGST)基因cDNA全序列与5′调控区的克隆与分析

淡水鱼类可溶性谷胱甘肽S-转移酶(sGST)在微囊藻毒素去毒代谢过程中具有独特的关键作用,因而也称为微囊藻毒素去毒酶.从淡水食毒藻鱼类鲢鱼(Hypophthalmichthysmolitrix)肝脏通过简并引物克隆微囊藻毒素去毒酶基因cDNA核心序列,应用5′RACE和3′RACE技术分别扩增该序列的5′末端和3′末端序列,最后通过序列拼接获得鲢鱼肝脏微囊藻毒素去毒酶基因cDNA全序列.序列分析结果表明,鲢鱼肝脏微囊藻毒素去毒酶基因cDNA全长920bp,其中5′-UTR长74bp,3′-UTR长174bp,编码区长672bp,编码223个氨基酸.应用基因组步行法,在鲢鱼克隆得到淡水鱼类微囊藻毒素去毒酶基因5′侧翼区878bp序列.与哺乳动物及海水鱼sGST基因不同,鲢鱼微囊藻毒素去毒酶基因的5′侧翼区,发现存在多个脂多糖反应元件(LPSRE),表明来源于毒藻的脂多糖可能对鲢鱼微囊藻毒素去毒酶基因表达有潜在调控作用.