小鼠15号染色体上脊髓重数量性状基因座的精细定位

目的以前的研究结果表明,控制小鼠脊髓重的一个数量性状基因座(QTL)位于15号染色体D15Mit158附近,跨度约30cM。为分离和确认脊髓重相关基因,本文对该QTL区域进行了精细定位。方法以高级互交系小鼠A/J×C57BL/6J(F4)为研究对象,选择脊髓重偏向两极的个体,在D15Mit158位点附近作高密度局部基因组扫描,用Map Manager QTX19软件对脊髓重与基因型进行连锁不平衡分析。结果在15号染色体D15Mit107附近出现了一个很强的连锁峰,LRS值为17.3(P=1.8×10-4),变异解释率为27%,LOD值达到3.75,可以认定为一主效QTL。该QTL跨度范围为3.2cM。另一个提示可能具有连锁关系的QTL位点在D15Mit28附近,LRS值为7.6(P=0.02),变异解释率为13%,跨度范围为5.0cM。结论控制小鼠脊髓重的D15Mit158区域实际上含有两个QTL,其中一个主效QTL位于15号染色体上宽约3.2cM的D15Mit107位点附近;另一个可能的QTL位于宽约5.0cM的D15Mit28附近。     

两种白斑小鼠突变基因的染色体定位

以本中心ENU诱变获得的两种白斑突变小鼠W-4Bao与Kitl-1Bao为研究对象[均为C57BL/6J(B6)背景],遗传试验表明它们都为单基因显性遗传,W-4Bao及Kitl-1Bao突变基因纯合子小鼠的表型分别为全白色及“黑头白”;将白斑杂合子小鼠与DBA/2(D2)交配获得具有白斑表型的F1小鼠,F1小鼠再回交D2繁殖[(B6×D2)F1×D2]F2小鼠,利用微卫星标记对F2代小鼠进行连锁分析。结果发现W-4Bao与微卫星D5Mit356、D5Mit308之间的LOD值分别为56.82、51.50,从而把该突变基因定位于第5号染色体D5Mit356与D5Mit308之间;Kitl-1Bao与微卫星D10Mit70、D10Mit68之间的LOD值分别为27.37、21.20,从而把该突变基因定位于第10号染色体上D10Mit70与D10Mit68之间。经过检索小鼠基因组数据库确认它们的候选基因分别为kit及kitl。     

两例新的稀毛小鼠突变基因的染色体定位

用连锁分析法对乙烷基亚硝基脲(ENU) 诱变获得的两例被毛突变小鼠(snthr 1Bao及snthr 2Bao) 的突变基因进行定位。选择平均分布于小鼠基因组且在C57BL/6J和DBA/2 间有差异的39 个微卫星对B6D2F1 互交得到的稀毛F2 进行全基因组扫描。扫描了9个微卫星后发现snthr 1Bao突变基因与D9Mit243 的LOD值为7 73。突变基因被定位于9号染色体。在此基础上又选择了D9Mit355 和D9Mit18 两个微卫星进行检测, 并扩大F2 的数量至145只。结果发现, snthr 1Bao与D9Mit18间无1 例重组, 稀毛突变基因与该微卫星紧密连锁, 距着丝点71cM。同理, 将snthr 2Bao突变基因也定位在与snthr 1Bao相近的区域。检索发现snthr 1Bao是一尚未克隆的新基因。     

ENU诱导获得一种短尾小鼠及其突变基因的初步定位

用一种化学诱变剂ENU(乙酰基亚硝基脲 )腹腔注射 3 0只 8～ 1 0周龄C5 7BL 6J(简称B6)雄鼠 (G0代 ) ,6周后与同品系正常母鼠配种繁殖后代 (G1代 )小鼠 3 5 1只。对其后代进行筛选获得一种可遗传的显性短尾突变小鼠。为了定位该突变基因 ,运用平均分布于B6和DBA 2 (简称D2 )小鼠常染色体而在这两者间又有差异的 3 9个微卫星对突变小鼠的 (D2×B6)F1代短尾突变小鼠回交D2得到的有短尾表型的[(B6×D2 )F1×D2 ]F2 代小鼠进行基因组扫描。反向运用经典的位置候选基因法 ,将短尾突变基因定位于 1 7号染色体 ,与D1 7Mit3 3的LOD值为 9 0 8。选用该染色体上与短尾表型相关基因Brachyury (T)最近的微卫星D1 7Mit1 43引物扩增 ,在 1 0 9只F2 代短尾小鼠中未发生一例交换 ,表明Brachyury基因是本例短尾突变强有力的候选基因。     

先天性骨—肾畸形综合征小鼠（KM—1d）致病基因ld的染色体定位

对ＫＭ－１ｄ小鼠的致病基因ｌｄ进行染色体定位,采用异构蛋白及同功酶电泳技术和体外扩增技术对同源导入近交系小鼠Ｃ５７ＢＬ／６．ＫＭ－ｌｄ２０对染色上的１４个笔化标记基因位点和６１个ＳＳＬＰ位点进行筛选,发现ｌｄ基因与２号染色体上的Ｄ２Ｍｉｔ３０、Ｄ２Ｍｉｔ６２和Ｄ２ｉｔ６３３个ＳＳＬＰ位点连锁,从而把ｌｄ基因初步定位于２号染色体,为进一步对ｌｄ基因准确定位,培育了８６只（Ｃ５７ＢＬ／６＊ＫＭ－１在＊     

一个新的小鼠被毛突变位点（Uncv）的高分辨遗传图谱和物理图谱的构建

迄今为止 ,人们已经发现了 10 0多个影响小鼠和人的毛发发育的基因 ,在以前的研究中 ,Uncv被证实是一个新的影响小鼠被毛的位点 ,具体表现为纯合突变体为无毛 ,杂合突变体表现为稀毛。除此之外 ,纯合的突变体还表现为生长和发育的迟缓。克隆这一突变基因将有助于更好地了解人类毛发发育异常相关的疾病。尽管这一位点已经被定位在小鼠 11号染色体上 ,但是没有该区域的精细遗传图谱和物理图谱直接克隆该基因有一定的难度。利用了两组杂交方式 ,[BALB/c(Uncv/Uncv)×C3H ( / ) ]×BALB/c (Uncv/Uncv)和 [BALB/c (Uncv/Uncv)×C5 7BL/6 ( / ) ]×BALB/c(Uncv/Uncv) ,共 2 0 74个F2代个体 ,通过对 11号染色体上的 16个微卫星标记的基因分型连锁分析 ,最终把该基因定位于11号染色体上位于D11Mit337和D11Mit338之间的约 1.4cM之间的区域。随后 ,利用BAC文库杂交和BAC末端序列PCR锚定的方法 ,构建了由 35个BAC构成的高分辨的BAC重叠群 ,这一高分辨的遗传图谱和物理图谱的构建 ,为进一步克隆这一突变基因打下了基础。     

PLCε基因敲除小鼠微卫星DNA遗传监测分析

目的利用多态性微卫星DNA位点分析PLCε基因敲除小鼠的遗传特性。方法用所筛选的15个微卫星DNA位点对28只PLCε基因敲除小鼠的DNA进行了PCR扩增,通过基因片段大小来分析群体的遗传多样性。结果 13个微卫星DNA位点中（D1Mit365、D3Mit51、D4Mit235、D6Mit102、D7Mit281、D8Mit113、D9Mit23、D10Mit180、D13Mit88、D16Mit145、D17Mit36、D18Mit94、D19Mit97）每个位点的28只小鼠DNA片段泳动距离一致,呈现单态性,表明该群体符合近交系的遗传特性;而利用Dq（敲基因型）和Dy（野生型）两个位点对28只小鼠的PCR扩增结果进行了鉴别分析,其中敲除基因型小鼠为6只;野生型为7只;杂合型为15只。结论利用微卫星标记技术可以对群体进行遗传质量监测,并能有效地鉴别不同的基因型,为小鼠的遗传质量监测提供了一种可行的方法。     

家蚕高密度AFLP连锁图谱的构建

为了进行家蚕Bombyx mori数量性状的QTL定位研究,以白色茧系品种C₁₀₀ (♀)和近交系大造(P50)(♂)杂交得到F₁,用F₁(♂)与双隐性标记的C₁₀₀ (♀)回交,得到回交一代(BC₁),用改进的AFLP分子标记方法,经96组选择性扩增引物扩增,获得分离比为1∶1(P≤0.05)的1 744个AFLP位点。用Map Manager QTXb19（Version 0.29）连锁图谱构建软件,构建了具有814个标记,36个连锁群的家蚕高密度AFLP分子标记连锁图谱。该连锁图谱覆盖的家蚕基因组长度为13 005 cM,连锁群长度变化范围为109.0～1 573.7 cM,连锁群的平均长度为361.25 cM,其标记间平均图距15.98 cM,最小图距2.3 cM,最大图距47.7 cM,标记间大于30 cM的gap共有39个。该连锁图平均每个连锁群23个标记,最多一个连锁群有92个标记,最少8个标记。该连锁图谱确定了与经典实验遗传图谱第15连锁群和W染色体连锁群相对应的两个连锁群。     

SNP标记对角膜混浊小鼠 突变相关基因的精细定位

为深入研究前期工作中以ENU诱变技术建立的遗传性角膜混浊突变系小鼠(B6-Co)的遗传机制, 利用 SNP标记对其突变基因进行精细定位, 将该品系中具有角膜混浊表型的小鼠(B6-CoP)与DBA/2小鼠(简称D2)配种得到F₁代, 再回交D2亲本品系得到F₂代, 提取F₂代角膜混浊小鼠鼠尾DNA。在MGI数据库中选取小鼠13号染色体已定位区间附近5个在C57BL/6(简称B6)和D2两个品系之间有差异的SNP, 应用聚合酶链反应-限制性片段长度多态性(PCR-RFLP)技术及连锁分析方法对B6-Co小鼠突变基因进行精细定位。结果表明: B6-Co小鼠突变基因定位于13号染色体上112 546 283~113 397 654 bp之间, 因该区间内有5个已知基因, 其中Map3k1基因与小鼠眼睛形态生成和眼睑闭合密切相关, 提示Map3k1是B6-Co小鼠突变的强力候选基因。     

小鼠39个微卫星的PCR条件及其运用

目的探索小鼠基因组39个微卫星的PCR条件,评价微卫星在小鼠遗传检测中的运用.方法采用梯度法探索39个微卫星的PCR条件;选择本中心不同来源及引种时间的C57BL/6、BALB/c、DBA/2J、CBA/N、FVB/NJ、ICR共6个品系(8个组)小鼠,每组采用10只个体的鼠尾,提取DNA并混合成DNA池,用39个微卫星扩增后电泳观察、比较种系纯度.结果小鼠微卫星的PCR条件差异较大,Mg2+浓度多数在1.5 mmol/L左右,退火温度多数在59℃左右.在6个品系小鼠的39个微卫星位点中,C57BL/6、BALB/c、DBA/2J都是纯合的; 其余品系有1～3个杂合位点. BALB/c在D5Mitl68、D8Mit320、D13Mit262三个位点,DBA/2J在D14Mit205位点与数据库记录有差异.结论本研究为小鼠39个微卫星提供了候选的PCR条件,并对6个品系小鼠的微卫星概貌及微卫星的运用价值进行了探讨.     

Recent advances in the study of Kaposi's sarcoma-associated herpesvirus replication and pathogenesis

It has now been over twenty years since a novel herpesviral genome was identified in Kaposi's sarcoma biopsies. Since then, the cumulative research effort by molecular biologists, virologists, clinicians, and epidemiologists alike has led to the extensive characterization of this tumor virus, Kaposi's sarcoma-associated herpesvirus(KSHV; also known as human herpesvirus 8(HHV-8)), and its associated diseases. Here we review the current knowledge of KSHV biology and pathogenesis, with a particular emphasis on new and exciting advances in the field of epigenetics. We also discuss the development and practicality of various cell culture and animal model systems to study KSHV replication and pathogenesis.     

A Unique Protease Cleavage Site Predicted in the Spike Protein of the Novel Pneumonia Coronavirus (2019-nCoV) Potentially Related to Viral Transmissibility

正Dear Editor,In December 2019, a novel human coronavirus caused an epidemic of severe pneumonia(Coronavirus Disease 2019,COVID-19) in Wuhan, Hubei, China(Wu et al. 2020; Zhu et al. 2020). So far, this virus has spread to all areas of China and even to other countries. The epidemic has caused 67,102 confirmed infections with 1526 fatal cases     

Curcuminoids as inhibitors of thioredoxin reductase: A receptor based pharmacophore study with distance mapping of the active site

Curcumin is the yellow pigment of turmeric that interacts irreversibly forming an adduct with thioredoxin reductase (TrxR), an enzyme responsible for redox control of cell and defence against oxidative stress. Docking at both the active sites of TrxR was performed to compare the potency of three naturally occurring curcuminoids, namely curcumin, demethoxy curcumin and bis-demethoxy curcumin. Results show that active sites of TrxR occur at the junction of E and F chains. Volume and area of both cavities is predicted. It has been concluded by distance mapping of the most active conformations that Se atom of catalytic residue SeCYS498, is at a distance of 3.56 from C13 of demethoxy curcumin at the E chain active site, whereas C13 carbon atom forms adduct with Se atom of SeCys 498. We report that at least one methoxy group in curcuminoids is necessary for interation with catalytic residues of thioredoxin. Pharmacophore of both active sites of the TrxR receptor for curcumin and demethoxy curcumin molecules has been drawn and proposed for design and synthesis of most probable potent antiproliferative synthetic drugs.     

The structure, reproduction and taxonomy of Vidalia and Osmundaria (Rhodophyta, Rhodomelaceae)

Comprises species occurring mostly in subtidal habitats in tropical, subtropical and warm-temperate areas of the world. An analysis of the type species, V. spiralis (Sonder) Lamouroux ex J. Agardh, a species from Australia, establishes basic characters for distinguishing species in the genus. These characters are (1) branching patterns of thalli, (2) flat blades that may be spiralled on their axis, (3) width of the blade, (4) primary or secondary derivation of sterile and fertile branchlets and (5) position of sterile and fertile branchlets on the thalli. Application of the latter two characters provides an important basic method for separation of species into three major groups. Osmundaria , a genus known only in southern Australia, was studied in relation to Vidalia , and its separation from the Vidalia assemblage is not accepted. Species of Vidalia therefore are transferred to the older genus name, Osmundaria. Two new species, Osmundaria papenfussii and Osmundaria oliveae are described from Natal. Confusion in the usage of the epithet, Vidalia fimbriala Brown ex Turner has been clarified, and Vidalia gregaria Falkenberg, described as an epiphyte on Osmundaria pro/ifera Lamouroux, is revealed to be young branches of the host, Osmundaria prolifera.     

New or rare chromosome counts in Artemisia L. (Asteraceae, Anthemideae) and related genera from Kazakhstan

Fifteen chromosome counts of six Artemisia taxa and one species of each of the genera Brachanthemum, Hippolytia, Kaschgaria, Lepidolopsis and Turaniphytum are reported from Kazakhstan. Three of them are new reports, two are not consistent with previous counts and the remainder are confirmations of very scarce (one to four) earlier records. All the populations studied have the same basic chromosome number, x = 9, with ploidy levels ranging from 2x to 6x. Some correlations between ploidy level, morphological characters and distribution are noted.