草坪型扁穗牛鞭草种质资源的坪用价值初步评价

采用景观-性能-应用适合度的综合评价体系,以暖季型狗牙根和冷季型高羊茅为对照,分春季、夏季、秋冬季对西南地区扁穗牛鞭草坪用性状和使用性能进行评价,结果表明：春季草坪景观质量H050、H049的坪用性状好于对照;夏季H036、H049、H011坪用性状好于对照;秋冬季H036、H049表现好于对照;综合一年的表现进行指标权重分析,H036、H049得分分别为17．60和18．08,可作为过渡性气候区建植观赏草坪、游憩草坪、运动草坪、保土草坪的优质材料进行开发利用。     

Resistance of SKW6 cell to apoptosis induction with anti-Fas antibody upon transduction of a reverse fragment to a cDNA encoding human 6A8 α-mannosidase

The effect of transduction with a reverse fragment to a cDNA encoding human 6A8 α-mannosidase on apoptosis induction of human B cell line SKW6 by anti-Fas antibody was tested. Apoptosis-inducer of anti-Fas monoclonal antibody was used to induce apoptosis in SKW6 cells. Giemsa’s staining, Annexin-V-FLUOS staining and DNA ladder test were used to determine the events of apoptosis. Indirect immunofluorescent staining with anti-Fas antibody was performed to detect the surface Fas expression. In a time-course test of 12, 24 and 36 h for apoptosis induction by anti-Fas antibody, DNA ladder was observed in the wild-type SKW6 cells in a time-dependent fashion. Mock transduction had no effect on DNA ladder production. However, no DNA ladder was detected in the rAAV-antisense 6A8 cDNA-transduced SKW6. Results from Annexin-V-FLUOS staining on anti-Fas antibody-treated cells revealed that the staining-positive rate in the rAAV-antisense 6A8 cDNA-transduced SKW6 cells was decreased in comparison to that in the wild-type and the mock-transduced cells. Giemsa’s staining observation showed that the number of dying (with apoptotic bodies) and dead cells was reduced in the rAAV-antisense 6A8 cDNA-transduced SKW6 cells in comparison with that in the wild-type and the mock-transduced cells upon anti-Fas antibody induction. The transduction did not affect the expression of Fas molecular on cell surface. 100% cells in all the groups showed Fas expression. The SKW6 cells became resistant to apoptosis induction by anti-Fas antibody upon transduction with a reverse fragment to a cDNA encoding human 6A8 α-mannosidase. The transduction did not affect the expression of Fas molecule on cells.     

Cloning and characterization of a cDNA encoding human differentiation antigen 5D4

A 1 846 bp cDNA is isolated from a human tonsil cell λgt 11 cDNA library (ATCC No. 37546) with mAb 5D4 reactive strongly with human B cell line 3D5, but weakly with human B cell line Daudi and human T cell line Jurkat as a probe. RT-PCR also shows a strong reaction in 3D5 cell and a weak reaction in Daudi and Jurkat cell for 5D4 mRNA. There is an open reading frame from 88 to 1 209 bp in 5D4 cDNA encoding a 374 AA protein. Both the Northern blot analysis and the two consecutive stop codens before start coden demonstrate that the cDNA is a full-length cDNA. Secondary structure prediction suggests that there are a region from 295 to 334 AA in the protein with strong hydrophobicity and a transmembrane helix region with high score from 313 to 334 AA with an orientation from the inside to the outside of the cell.     

野生扁穗牛鞭草无性系构件组成及生物量结构变异性

对60份野生扁穗牛鞭草的无性系种群构件数量、质量性状及生物量结构进行比较分析,结果说明：不同种群构件性状与数量呈现出变异性,叶长、叶宽、单蘖叶片数、无性系叶片数的变异系数分别为24．95％、20．00％、14．12％、43．56％;茎直径、节直径、节间长、单蘖节数、直立茎长度、匍匐茎长度、直立茎数、匍匐茎数的变异系数分别为21．11％、22．42％、20．10％、12．14％、46．73％、20．14％、72．76％、37．97％;根系深度、根系分布范围、分蘖面积的变异系数分别为19．81％、37．65％、33．68％;花序长、花序宽、花序厚、单蘖花序数、直立茎生殖蘖比例、匍匐茎生殖蘖比例的变异系数分别为13．33％、13．42％、14．80％、36.10％、118．96％、81．44％。不同种群构件生物量结构变异性丰富,叶生物量、茎生物量、根生物量、花序生物量变异系数分别为78．51％、91．66％、45．64％、192．24％;构件的数量差异高于性状差异,无性系种群差异主要体现在分蘖能力与空间拓展能力上。野生资源生态型变异性和对环境的适应性,为优异性状选择、品种选育、资源开发利用提供了丰富的物质基础。     

SKW6细胞在转导6A8 a-甘露糖苷酶反义cDNA后对抗Fas抗体诱导的凋亡

鉴于蛋白质糖基化的重要生物学意义, 以腺相关病毒为载体, 把编码6A8 a-甘露糖苷酶的cDNA 3′端1358 bp的反向片段转导入EB病毒转化的B细胞株SKW6, 探讨6A8 a-甘露糖苷酶表达抑制对抗Fas抗体诱导凋亡的影响. 反义6A8转导成功及表达用Northern杂交及RT-PCR检测, 6A8 a-甘露糖苷酶表达用Con A结合试验检测. Giemsa染色, AnnexinⅤ染色及梯形DNA电泳显示, Fas抗体能诱导SKW6细胞凋亡, 但6A8 a-甘露糖苷酶表达抑制的细胞对Fas抗体诱导的凋亡发生抵抗, 而转导正义6A8或空载载体则无影响. 6A8 a-甘露糖苷酶表达状况对SKW6细胞表面Fas分子表达没有影响.     

编码人分化抗原5D4的全长cDNA克隆及其编码蛋白质的二级结构分析

用 3D5细胞染色强阳性 ,而Daudi与Jurkat细胞染色弱阳性的单克隆抗体5D4,从一个人扁桃体细胞λgt1 1cDNA文库 (ATCCNo .375 46 )筛选到一个长 1 846bp的cDNA克隆 .RT_PCR显示 3D5细胞、Daudi细胞和Jurkat细胞 5D4mRNA均阳性 ,但 3D5细胞明显强于后两者 . 5D4cDNA中 88～ 1 2 0 9bp为一个开放阅读框架 ,编码374个氨基酸的蛋白质 .Northernblot分析结果和起始密码子前 5′端非编码区有 2个连续的终止密码子表明 ,这是一个全长cDNA .从其开放阅读框架推断的蛋白质的氨基酸序列中 2 95～ 334aa为一强疏水区 ,预测 31 3～ 334aa为一分值极高的跨膜螺旋区 ,其方向最可能是由膜内到膜外 .     

Resistance of SKW6 cell to apoptosis induction with anti-Fas antibody upon transduction of a reverse fragment to a cDNA encoding human 6A8 α-mannosidase

The effect of transduction with a reverse fragment to a cDNA encoding human 6A8 α-mannosidase on apoptosis induction of human B cell line SKW6 by anti-Fas antibody was tested. Apoptosis-inducer of anti-Fas monoclonal antibody was used to induce apoptosis in SKW6 cells. Giemsa's staining, Annexin-V-FLUOS staining and DNA ladder test were used to determine the events of apoptosis. Indirect immunofluorescent staining with anti-Fas antibody was performed to detect the surface Fas expression. In a time-course test of 12, 24 and 36 h for apoptosis induction by anti-Fas antibody, DNA ladder was observed in the wild-type SKW6 cells in a time-dependent fashion. Mock transduction had no effect on DNA ladder production. However, no DNA ladder was detected in the rAAV-antisense 6A8 cDNA-transduced SKW6. Results from Annexin-V-FLUOS staining on anti-Fas antibody-treated cells revealed that the staining-positive rate in the rAAV-antisense 6A8 cDNA-transduced SKW6 cells was decreased in comparison to that in the wild-     

Lnc-RAP3对小鼠3T3-L1前脂肪细胞分化的影响

长链非编码RNA(long non-coding RNA,lnc RNA)是一类长度大于200nt、没有长开放阅读框架但往往具有m RNA结构特征的RNA,可以在转录及转录后水平参与基因的表达调控。近年来,有研究证实lnc RNA对脂肪生成具有重要作用。Lnc-RAP3位于小鼠(Mus musculus)17号染色体,其表达量在小鼠脂肪细胞分化前后呈现显著差异,但其具体的生物学功能尚不清楚。为探讨lnc-RAP3在小鼠3T3-L1前脂肪细胞成脂分化中的作用,本文首先构建了lnc-RAP3的真核表达载体pc DNA3.1-RAP3,利用脂质体将pc DNA3.1-RAP3和人工合成的lnc-RAP3的si RNAs分别转染3T3-L1前脂肪细胞,并对转染后的细胞进行诱导分化,并通过油红O染色、q RT-PCR检测成脂分化相关基因表达等方法比较过表达和敲降lnc-RAP3对3T3-L1前脂肪细胞成脂分化的影响。结果显示,过表达lnc-RAP3后,细胞内脂滴聚集显著减少(P0.05),在诱导分化第0 d、2 d和4 d时C/EBPα、Glut4、PPARγ、LPL和FAS的表达水平均呈显著(P0.05)或极显著(P0.01)下降;敲降lnc-RAP3后,细胞内脂滴聚集显著增多(P0.05),同时在诱导分化第0 d、2 d时PPARγ、LPL、C/EBPα、FAS和Glut4的表达水平呈显著(P0.05)或极显著(P0.01)升高。本研究结果表明,lnc-RAP3可能通过影响成脂分化相关基因的表达来抑制3T3-L1前脂肪细胞的成脂分化。     

CRISPR-Cas9技术的原理及其在猪研究中的应用

成簇的规律间隔的短回文重复序列(clustered regularly interspaced short palindromic repeats,CRISPR)及其相关蛋白(CRISPR-associated proteins,Cas)是细菌和古生菌中抵抗外源病毒或质粒入侵的获得性免疫系统,目前已经被广泛应用于动物基因组编辑。现回顾CRISPR-Cas9系统的发展历程并比较其与锌指核酸酶(ZFNs)、类转录激活因子效应物核酸酶(TALENs)技术的优势,详细介绍了CRISPR-Cas9系统的组成成分和各组分的功能以及其编辑基因组的原理,着重梳理了CRISPR-Cas9系统在猪生产性能、抗病育种、人类模式动物构建和异种器官移植方面的最新研究进展,以期为CRISPR-Cas9系统的进一步应用提供参考。     

Inhibition of metastasis to lung of a human nasopharyngeal carcinoma cell line CNE-2L2 transfected with pRc/CMV-antisense 6A8 cDNA in nude mice

The growth of CNE-2L2 cell, a cloned line of human nasopharyngeal carcinoma with a high potentiality of metastasis to lung was inhibited to a certain extent after transfection with a recombinant antisense expression vector of a cDNA encoding a human α-mannosidase (pRc/CMV-antisense 6A8 cDNA)( the Genbank accession number of 6A8 cDNA is U37248) in comparison with that of the cell transfected with the Mock and of the wild cell. Two months after a subcutaneous inoculation of CNE-2L2 cell into the axilla of nude mice metastatic lesions in the lung were observed in 9/10 mice (90%) with grade Ⅲ in 8 mice and grade Ⅱ in one mouse in the wild cell group, in 6/8 mice (75%) with grade Ⅲ in one mouse, grade Ⅱ in 2 mice and grade Ⅰ in 3 mice in the Mock-transfection group, in only 3/10 mice (30%) with all grade Ⅰ in pRc/CMV-antisense 6A8 cDNA-transfection group.