Modification of alternative splicing in bovine somatic cell nuclear transfer embryos using engineered CRISPR-Cas13d

Animal cloning can be achieved by somatic cell nuclear transfer(SCNT), but the resulting live birth rate is relatively low. We previously improved the efficiency of bovine SCNT by exogenous melatonin treatment or by overexpression of lysine-specific demethylase 4D(KDM4D) and 4E(KDM4E). In this study, we revealed abundant alternative splicing(AS) transitions during fertilization and embryonic genome activation, and demonstrated abnormal AS in bovine SCNT embryos compared with in vitro fertilized ...     

中国东部主要松林营养元素循环的比较研究

 中国的松林主要分布在亚热带和温带地区,在亚热带和温带地区东部主要是马尾松林、华山松林、油松林、红松林和樟子松林。松林由于树种、起源和年龄的差别,其生物量的变化幅度较大,在65～200t·hm-2之间(东北地区的原始红松林最高生物量可达360t·hm-2),松林的生物量表现出区域分异的特点。即从南到北随着纬度的增加,林分的生物量有逐渐降低的趋势。松树针叶中5种主要营养元素含量表现为[N]>[K]≥[Ca]>[Mg]≥[P],而且营养元素表现出因种而异,N的含量为华山松≥马尾松>油松≥红松>樟子松,而P和K在油松和红松针叶中含量较高;Ca的含量表现出较大的波动,与其母岩关系密切。松林主要营养元素(N、P、K、Ca、Mg)的积累量中N一般占25％～40％,松林营养元素循环速率受生境,树种、年龄的影响,但总的来说,亚热带地区松林营养元素的循环速率高于温带地区松林。     

多效唑(PP333)对美洲商陆毛状根生长和商陆皂苷甲产生的影响

为了探究植物生长调节剂多效唑(PP333)调控药用植物美洲商陆(Phytolacca americana)毛状根生长和次生代谢的可能性, 设计实验并探讨PP333对美洲商陆毛状根生长及其商陆皂苷甲含量的影响。结果表明, 与对照相比, PP333使毛状根根尖及侧根表面呈浅红色, 侧根变得短而粗, 且随着培养基内PP333浓度的升高, 根表面颜色加深。培养基中添加0.5-5.0 mg·L^-1 PP333能促进毛状根中商陆皂苷甲的产生, 其中以1.0 mg·L^-1 PP333的效果最好, 其商陆皂苷甲含量达6.22 mg·g^-1 DW, 约为对照的1.94倍。PP333能提高毛状根苯丙氨酸裂解酶(PAL)的活性, 并可能通过对PAL酶活性的调节来促进毛状根中商陆皂苷甲的产生。     

水稻大粒种质资源和遗传分析

谷粒重量是构成产量的三要素之一, 对提高水稻产量具有重要意义。本文概述了国内外水稻大粒种质资源的现状, 同时对粒重基因遗传分析的研究进展进行了综述。粒重是一个受多基因控制的数量性状, 目前定位的粒重数量性状位点至少达89个、精细定位1个粒重基因gw3.1和１个长粒基因Lk-4(t)以及克隆1个粒重基因GS3, 并在此基础上讨论了粒重在育种上的应用。     

絮凝基因的克隆和在工业啤酒酵母菌株中表达

The partial genomic library of Saccharomyce cerevisiae FL189 possessing strong flocculation ability was constructed using Yeast-E.coli shuttle plasmid YCp50 as vector.Recombinant plasmid containing flocculation gene was obtained by screening the growth of transformants on the selective medium and measurement flocculation,designated as pCF1.pCF1 was introduced into industrial yeast strain PJ208-5-15.Six transformants PJ208-5-15-1(pCF1)～PJ208-5-15-6(pCF1)possessing strong flocculation ability were obtained.Th…     

几种药物抑制马传染性贫血病毒和人免疫缺陷病毒的实验研究

用马传染性贫血病毒—驴胚肺二倍体细胞(EIAV-DELDC)为实验体系,以细胞中病毒逆转录酶活性及病毒相关抗原的表达为观察指标,检测了叠氮胸苷(AZT)、三氮唑核苷(Ribavirin,病毒唑)、磷羧基甲酸钠(PFA)和苏拉明等4种已知抗人免疫缺陷病毒(HIV-1)药物对马传染性贫血病毒的抑制作用。结果表明,PFA、AZTTP(三磷酸AZT)和苏拉明均能抑制病毒相关抗原的表达,AZT虽无此作用,但能抑制细胞内逆转录酶活性。用~3H-TMP掺入法比较了PFA、AZTTP、苏拉明对体外无细胞系EIAV逆转录酶粗提物和HIV-1基因工程产物逆转录酶活性的抑制作用表明,两种逆转录酶对苏拉明的敏感性相近,而HIV-1逆转录酶对PFA和AZTTP的敏感性较EIAV者高约100倍。又以无细胞系中逆转录酶活性测定法,检测了12种中药提取物的抑制作用,其中小柴胡汤对EIAV和HIV-1逆转录酶活性都有抑制作用,IC_(50)为717μg/ml和700μg/ml(生药浓度)。小柴胡汤对两种病毒感染细胞中抗原的表达和HIV引起细胞病变都有抑制作用,对HIV-1的抑制比EIAV强。这些结果表明,EIAV-DELDC体系可考虑作为抗HIV-1药物筛选模型。     

Pull-push neuromodulation of LTP and LTD enables bidirectional experience-induced synaptic scaling in visual cortex

Neuromodulatory input, acting on G protein-coupled receptors, is essential for the induction of experience-dependent cortical plasticity. Here we report that G-coupled receptors in layer II/III of visual cortex control the polarity of synaptic plasticity through a pull-push regulation of LTP and LTD. In slices, receptors coupled to Gs promote LTP while suppressing LTD; conversely, receptors coupled to Gq11 promote LTD and suppress LTP. In vivo, the selective stimulation of Gs- or Gq11-coupled receptors brings the cortex into LTP-only or LTD-only states, which allows the potentiation or depression of targeted synapses with visual stimulation. The pull-push regulation of LTP/LTD occurs via direct control of the synaptic plasticity machinery and it is independent of changes in NMDAR activation or neuronal excitability. We propose these simple rules governing the pull-push control of LTP/LTD form a general metaplasticity mechanism that may contribute to neuromodulation of plasticity in other cortical circuits.     

siRNA binding proteins of microglial cells: PKR is an unanticipated ligand

Small interfering RNA (siRNA), double-stranded RNA (dsRNA) 21-23 nucleotides (nt) long with two nt 3' overhangs, has been shown to mediate powerful sequence-specific gene silence in mammalian cells through RNA interference (RNAi). Due to its high efficiency and high specificity siRNA has been used as a powerful post genomic tool and a potent therapeutic candidate. However, there is still a lot to learn about the mobility of siRNA inside cells and the cellular factors that might interfere with the specificity and activity of siRNA. Microglia are the brain's effector cells of the innate immune system and suitable targets in the development of novel therapeutic strategies. Here, we show the cellular uptake and intracellular distribution of siRNA in murine microglial N9 cells. siRNA was internalized by microglial N9 cells without transfection reagent and mainly localized to the endosomes However, no significant gene silencing effects were observed. Its cellular uptake and cellular distribution pattern were similar with that of a same length single stranded DNA (ssDNA). Further, cellular binding proteins of siRNA were purified and identified by mass spectrometry. Negative control siRNA and siRNA targeted to beta-actin were used in this part of experiment. Most of the siRNA binding proteins for negative control siRNA and siRNA targeted to beta-actin were dsRNA-binding proteins, such as dsRNA-dependent protein kinase R (PKR). Furthermore, both control siRNA and siRNA targeted to beta-actin activated PKR in N9 cells, which suggest that siRNA might cause off-target effects through activation of PKR.     

ROCK1 induces ERK nuclear translocation in PDGF-BB-stimulated migration of rat vascular smooth muscle cells

It has been known that Rho-associated protein kinase (ROCK) signaling regulates the migration of vascular smooth muscle cells (VSMCs). However, the isoform-specific roles of ROCK and its underlying mechanism in VSMC migration are not well understood. The current study thus aimed to investigate the roles of ROCK1/2 and their relationship to the MAPK signaling pathway in platelet-derived growth factor (PDGF)-induced rat aorta VSMC migration by manipulating ROCK gene expression. The results revealed that ROCK1 small interfering ribonucleic acid (siRNA) rather than ROCK2 siRNA decreased PDGF-BB-generated VSMC migration, and upregulation of ROCK1 expression via transfection of constructed pEGFP-C1/ROCK1 plasmid further increased the migration of PDGF-BB-treated VSMCs. In PDGF-treated VSMCs, ROCK1 siRNA did not affect the phosphorylation levels of ERK and p38 in the cytoplasm, but decreased the level of ERK phosphorylation in the nucleus. These findings demonstrate that activated ROCK1 can promote VSMC migration through facilitating phosphorylation and nuclear translocation of ERK protein.