鸡Myostatin基因单核苷酸多态性的群体遗传学分析

肌肉生长抑制素是控制骨骼肌生长发育的重要细胞因子,采用PCR－SSCP和测序的方法发现了5个位于Myostatin基因5′－和3′－调控区的单核苷酸多态性位点,对北京油鸡、白耳鸡、石歧杂、矮小黄鸡、小型黄鸡、惠阳胡须鸡、隐性白羽鸡、海兰、AA鸡等不同鸡种的该单核苷酸多态性分析结果表明：Myostatin基因的5′调控区引物P60／P61扩增片段多态性是由3个核苷酸的改变而产生的[分别是G→A（304位）、A→G（322位）、G→（344位）],引物P93／P94扩增片段的多态性是由G→A（167位）突变造成的,引物P117。P118PC扩增片段多态性是由T→C（177位）造成的。3′调控我引物P80／P81扩增片段多态性是由第7263位A突变为T造成的,引物P76／P77扩增片段多态性是由A→G（6935位）造成的。不同鸡种群体遗传学分析表明,5′－调控区引物60／P61扩增片段多态性片段多态性是由A→G（6935位）造成的。不同鸡种群体遗传学分析表明,5′－调控区引物P60／P61扩增片段多态性位点在北京油鸡的基因型频率分布与其他的品种有很大的差异,其BB型频率为0．700,AA基因型频率仅为0．033,而其他鸡种中以A基因优势;对于引物P93／P94,品种间的基因型频率差异极显著（P＜0．01）,北京油鸡和AA鸡的EE型频率鸡种中以A基因占优势;对于引物P93／P94,品种间的基因型频率差异极显著（P＜0．01）,北京油鸡和AA鸡的EE型频率低于其他品种,白耳鸡和海兰蛋鸡以EE型为主,其频率高于其他品种;3′－调控区引物P80／P81多态怀位点在9个鸡种中都是等位基因C占优势。引物P76／P77,总体上MM型的频率较低,杂合子MN型的频率较高。     

巨型引物PCR法对抗CD3改型单链抗体

亲和力是影响改型单链抗体应用于临床的重要因素之一.利用巨型引物PCR定点诱变方法,设计并化学合成出两组含多个突变位点的简并引物,在第一轮PCR中使用简并引物分别扩增出含突变碱基的两条特异性的DNA片段,即巨型引物,将其经琼脂糖凝胶电泳分离纯化后,作为3′和5′的两端引物应用于第二轮PCR反应中.通过改变标准PCR反应条件,调整引物与模板的浓度,扩增出特异性较强的目的DNA条带.PCR产物经回收后,进行DNA测序.测序结果表明利用该方法扩增得到特异的抗CD3改型单链抗体的突变体库.     

油菜单显性核雄性不育基因的分子标记

以陕西省杂交油菜研究中心选育的单显性核不育油菜分离群体为材料,利用集群分离法（BSA）对该油菜单显性核不育基因进行了RAPD分析。在随机选取的300个10碱基随机引物中,引物S243（5′CTATGCCGAC3′）在可育集团与不育集团间扩增出特异而可重复的1．5kb的多态性片段OPU-031500,而在细胞质雄性不育和其它核不育类型油菜中均未扩增出上述特异性片段,从而确证此RAPD标记OPU-031500。片段是与甘蓝型油菜单显性核不育基因连锁的。将该多态性片段克隆并测序,发现其序列与拟南芥的一段DNA序列高度同源。根据同源序列及测序结果设计两对特异引物（P1／P2和P3／P4）,引物P3／P4在可育系中可扩增到约1．5kb的单一特异片断,而在不育系中无带,从而将RAPD标记转化为稳定可靠的SCAR标记。     

封闭群斑马鱼的STR遗传检测技术

目的以AB、LF、ST（本地短尾,short tail）三个封闭群斑马鱼的基因组DNA为主要研究对象,采用STR（short tandem repeat）引物扩增方法分析不同种群在核酸水平的异同。方法利用5对斑马鱼STR引物和3对剑尾鱼STR引物,优化PCR条件,对不同种群斑马鱼DNA进行扩增。结果Z24、Z9384、Z10508、Z20046这4对引物可扩增出稳定条带。除Z24的3对STR引物在群内和群间均表现出多态性。对扩增不同带型进行统计分析,Z20046群间差异显著（P〈0.01）,Z9384有群间差异（P〈0.05）,Z10508扩增结果无统计学差异（P〉0.05）。结论鉴于该方法操作简便,通过进一步研究,可为封闭群斑马鱼的遗传质量评价打下基础。     

洋葱细胞质雄性不育系70及其保持系71基因组DNA的RAPD分析

采用RAPD技术,对洋葱细胞质雄性不育系70及其保持系71的基因组DNA的多态性进行分析。结果表明,在被检测的100条随机引物中,46条引物具有扩增产物,5条引物扩增出多态性条带。对这5条引物进行重复筛选和单株验证,只有G02扩增出的多态性片段,且表现稳定。推测该标记与育性有关。     

一步法定点突变技术快速构建bsh基因突变启动子

目的：建立一种高效便捷的定点突变方法,为基因表达调控以及蛋白质结构和功能的研究提供技术支撑。方法：以构建单核细胞增生李斯特菌（Listeria monocytogenes）中编码胆碱水解酶（bile salt hydrolase,BSH）的bsh基因突变启动子为例,采用一对完全互补并带有突变位点的引物扩增携带bsh基因启动子的重组质粒DNA全序列,通过DpnⅠ消化PCR产物中剩余的甲基化的模板DNA,酶切后的PCR产物直接转化大肠杆菌,从而获得含有突变启动子的重组质粒。结果：通过一步法定点突变技术成功构建了bsh基因的三种突变启动子。结论：该方法简单高效,只要把握好对引物设计,高保真的DNA聚合酶、模板DNA的浓度以及PCR扩增程序的选择,突变成功率可以达到100%。     

玉米同核同质系在研究细胞质雄性不育中的应用

以94个Operon公司的10碱基RAPD引物,对玉米同核同质系（I）中的野生型正常可育株478（F）和其突变的不育株478－cms的线粒体DNA(mtDNA)进行了RAPD分析。结果表明：仅OPZ－19、OPAA－15与OPS－01 3个引物在同核同质系（I）的野生型正常可育株和其突变的不育株之间扩增出多态产物OPZ－19420、OPAA－15600与OPS－1400。提示在同核同质系的野生型正常可育株和其突变的不育株的mtDNA之间同源性很高,多态性很低,同核同质系相当于在mtDNA水平上的近等基因关系,所找到的多态就在所研究的目标基因附近,因此它是研究mtDNA基因的良好体系。上述3个多态扩增结果在同核同质体系（I）的姐妹同核同质系（Ⅱ）中进行了验证, 结果表明仅2个引物OPZ－19、OPAA－15在姐妹同核同质系（Ⅱ）的野生型正常可育株苏478（F）和其突变的不育株苏478－cms之间获得了与同核同质系（Ⅰ）相同的多态性扩增产物OPZ－19420、OPAA－15600。即这两个多态性产物在两套同核同质系中同时存在,因而这两个多态性产物与育性直接相关的可能性很大。     

甜菜银叶病菌的PCR检测

本研究用16S23S rDNA间的ITS 序列通用引物L1（5′AGTCGTAACAAGGTAGCCGT3′）和L2 （5′ GTGCCAAGGCATCCACC3）扩增甜菜银叶病菌（Curtobacterium flaccumfaciens pv. betae,Cfb）和其它相近细菌的基因组DNA;并对其PCR产物进行回收、克隆和测序,将所获序列和其它已报道的细菌内源转录间隔区(Internally Transcribed Spacer,ITS)序列进行多重比较后设计出Cfb的特异性引物B1（5′GGCCTCGTGTTGTCCCTTATC3′）和B2 （5′GTCACCAATCAACAACCCGAG3′）。此引物可以从Cfb中扩增出387bp 的特异性片段,而其余参试的21个细菌PCR反应结果均为阴性。该方法可以应用于病害防治工作中的Cfb快速、可靠的检测。     

RAPD法对罗曼蛋鸡基因组DNA多态性分析初报

利用RAPD（随机扩增多态DNA）方法,选用37种10bp的随机引物,对罗曼蛋鸡基因组DNA进行多态性分析,发现其中两种引物（OPS－08,OPY－06）在三代7个品系中都能扩增出多条带并且反映其基因组DNA的多态性,可用于检测出不同品系间的差异,并且这两种引物的碱基序列有80％是对应互补的。     

体细胞克隆山羊微卫星DNA分析

用10对山羊微卫星DNA多态性引物对2只体细胞克隆济宁青山羊、青山羊供体细胞、受体奶山羊母羊以及具有亲缘关系的3只对照济宁青山羊进行微卫星DNA分析.结果表明有5对山羊微卫星DNA多态性引物,即SR-CRSP1, SR-CRSP5, SR-CRSP6, SR-CRSP7和SR-CRSP24,扩增产物有明显的多态性.扩增产物经过6%变性聚丙烯酰胺凝胶电泳后银染,结果2只体细胞克隆山羊的微卫星DNA指纹与供体细胞完全相同,而且不同于其受体母亲也不同于其他所有同品种不同个体的对照青山羊.证明体细胞克隆山羊基因组来源于供体细胞.     

六种藓类植物茎的比较解剖学研究

通过对6种藓类植物,即褶叶青藓(Brachythecium salebrosum(Web.et Mohr.)B.S.G.)、湿地匐灯藓(Plagiomnium acutum(Lindb.)Kop.)、侧枝匐灯藓(Plagiomnium maximoviczii(Lindb.)Kop.)、大凤尾藓(Fissidensnobilis Griff.)、大羽藓(Thuidium cymbifolium(Doz.et Molk.)B.S.G.)和大灰藓(Hypnum plumaeforme Wils.)嫩茎和老茎的石蜡切片和显微观察发现,同一藓类植株的嫩茎和老茎,茎结构稳定,不同种藓类植物茎横切面具有不同特征.植物体茎横切面形状、表层细胞的层数、细胞大小和细胞壁厚薄、皮层细胞大小和形状、中轴的有无以及比例等特征可以作为藓类植物的分科分类依据之一.     

Photoregulation of seed germination of wild-type and of an aurea-mutant of tomato

Seed germination of an aurea mutant of tomato ( Lycopersicon esculentum Mill.) is promoted by continuous irradiation with red, far-red or long-wavelength far-red (758 nm) light as well as by cyclic irradiations (5 min red or 5 min far-red/25 min darkness). Far-red light applied immediately after each red does not change the germination behaviour. Seed germination of the isogenic wild-type, cv. UC-105, is promoted by continuous and cyclic red light while it is inhibited by continuous and cyclic far-red light and by continious 758 nm irradiation. Far-red irradiation reverses almost completely the promoting effect of red light. The promoting effect (in the aurea mutant) and the inhibitory effect (in the wild-type) of continuous far-red light do not show photon fluence rate dependency above 20 nmol m⁻² s⁻¹. It is concluded that phytochrome controls tomato seed germination throgh low energy responses in both the wild type and the au mutant. The promoting effect of continuous and cyclic far-red light in the au mutant can be attributed to a greater sensitivity to P_fr.     

真菌类遗传学分析的知识结构教学

本文以认知结构理论为指导,讨论了真菌类遗传分析与高等动植物遗传分析的内在联系,认为利用这种内在联系进行教学可收到好的效果并说明了作者的具体教学过程。 Abstract:In the paper, the relationship between genetic analysis of Fungi and genetic analysis of high animal and plant was discussed.A good results were obtained when we adopted this method in the teaching.     

Cause of Senescence of Nine Sorts of Flowers

The levels of endogenous phytohormones and respiratory rate in nine sorts of flowers such as Cymbidium faberi Rolfe, Nopalxochia ackermannii Kunth and others were investigated both at full bloom and senescence and meanwhile the effect of exogenous phytohormones on prolonging the blossoms and promoting ethylene production were tested. There is a high content of endogenous ethylene in all the long-lived flowere, about 3–16 folds higer than the short-lived ones. There is a high level of ABA at full blooming flowers of short-lived flowers, in which there is no or only some cytokinins in it, but the ratio of CTK (6BA+zeatin)/ABA is smaller(l.7). The endogenous ABA reached a much higher level at senescence in all nine sorts of flowers, so it is reasonable to consider that it is ABA which plays an important role of regulation in controlling flower's senescence. There is a much higher level of GA3 and zeatin in the long-lived flowers which is not demonstrated in the shortlived ones. The respiratory rate is one of the factors controtling the longevity of flowers, but it does not play a decided role. Application of 6BA and zeatin prolongs distinctly orchid’s longevity, however exogenous IAA through the promotive action on ethylene production, evidently extends the longevity of the flowers of the Nopalxochia ackermannii Kunth.     

龙胆科药用植物化学成分的研究现状

龙胆科植物在我国的分布范围很广,且多数为药用植物,其多数种属的药用植物,至今其化学成分尚未被系统研究。综述了目前龙胆科药用植物的化学成分的研究现状及一般提取方法,对近年来发现的环烯醚萜及裂环烯醚萜类化合物进行了总结,为本科药用植物的更深入研究提供了参考。     

Interconnection of parameters of regulation system of lipid peroxidation and of morphophysiological parameters of mouse liver

A complex analysis of seasonal fluctuations of the mean group parameters of the system of regulation of lipid peroxidation has been performed in liver of Balb/c mice. Association of lipid characteristics and morphophysiological parameters is studied in the Balb/c mouse liver. An inter-connection is revealed between the liver index and the amount of lysoforms of phospholipids, the scale and character of the interconnection differing essentially depending on proportion of phos-phatidylcholine in mouse liver phospholipids.     

Time of detection of recessive genes: effects of system of mating and number of examined individuals

Summary Anthers were cultured from two sets of seven lines of hexaploid wheat (Triticum aestivum L.) with different cytoplasms, the euplasmic nucleus donors, Siete Cerros 66 and Penjamo 62, as well as their six alloplasmic lines derived from wild relative species of the genera Triticum and Aegilops. Significant cytoplasmic and nuclear effects but no cytoplasmic-nuclear interaction were found for embryogenic anther response, with the best performance of Penjamo 62 in Ae. kotschyi cytoplasm. Plant regeneration was not affected significantly by the cytoplasmic background of the lines cultured. The possible genetic implications of the observed cytoplasmic and nuclear influences on the in vitro haploid induction of wheat are discussed.