基因治疗及其在兽医学中的应用

基因治疗是20世纪90年代形成的，该技术的研究一开始就与动物医学的发展密不可分，主要是通过建立动物疾病模型分析和研究基因治疗各种层面上的问题，但它真正在兽医临床上的研究是近年才开始的。许多试验结果表明，一旦载体的安全性有了保障，基因治疗将有可能成为动物重大疫病有效预防和治疗的方法，在兽医学中有很广阔的应用前景。     

利用 ｍｔＤＮＡＤ-ｌｏｏｐ序列为遗传标记分析槟榔江水牛的群体遗传特征

槟榔江水牛是近年在云南西部发现的中国第一个本土河流型水牛群体,具有较高种用价值,但其重要遗传背景信息还不清楚。本文采用 ＰＣＲ产物直接测序法对 ８６头槟榔江水牛 ｍｔＤＮＡＤ-Ｌｏｏｐ序列进行了突变检测,并以 ＧｅｎＢａｎｋ上已发表的 ７０条河流型和 １１２条沼泽型水牛 ｍｔＤＮＡＤ Ｌｏｏｐ序列为对照,对所得数据进行群体遗传和系统发育分析。结果在槟榔江水牛中检测到 ３３种单倍型,１１２个多态位点。其中单一变异位点 １４个,简约信息位点 ９８个。槟榔江水牛 ｍｔＤＮＡＤ Ｌｏｏｐ序列 Ｔ,Ｃ,Ａ,Ｇ的平均含量分别为 ２８.４３％,２４.８２％,３１.９６％,１４.７９％,单倍型多样度为 （０.９４８±０.０１２）,核苷酸多样度为 （０.０３８１±０.００１６）,序列间平均核苷酸差异数为３３.２８８,群体内平均遗传距离为 （０.０４３±０.００５）。系统发育、中介网络图和群体遗传关系分析表明,槟榔江水牛含有两个差异显著的母系世系组分,其中一个为河流型世系,在群体中占 ６１.６３％;另一个为沼泽型世系,在群体中占３８３７％,而其沼泽型世系可进一步分为 Ａ,Ｂ,Ｃ３个支系,其中,Ｃ为在水牛中新发现的支系,其频率极低。结果揭示了槟榔江水牛群体遗传多样性丰富,但该群体存在一定的沼泽型水牛基因渗入。     

25份澳大利亚小麦种质的性状表现及利用

为给黄淮南片冬麦区小麦育种提供信息,对25份澳大利亚小麦种质于2016-2018年度在新乡进行了部分农艺性状调查,对其品质性状及优质亚基等进行了分析。结果表明,25份澳大利亚小麦种质多表现为春性,综合农艺性状较差;千粒重低于黄淮麦区高产对照品种周麦18,变化范围为31.4 g～43.1 g;湿面筋含量、面筋指数、稳定时间、弱化度的变化范围分别为28.0%～39.1%、55.2%～96.5%、4.0 min～25.0 min、5%～100%。从澳大利亚小麦种质中,筛选出了具有湿面筋含量高、面筋指数高、稳定时间长、面粉粉色白等优良品质特性的材料,鉴定出了含5+10亚基、7+8~*亚基(7超量表达亚基)、17+18亚基、13+16亚基、低PPO活性基因及 Wx-B1基因缺失等优异品质材料,筛选出了含有 Rht-D1b矮秆基因以及 Lr34和 Lr46抗叶锈基因的材料。利用澳大利亚小麦种质为改良亲本,与黄淮麦区表现突出的小麦品种配制三交或四交组合,按常规系谱方法对优良性状进行定向选择,培育出了综合农艺性状表现好、品质表现突出的小麦新品系。     

Nomenclature report on rice WRKY's - Conflict regarding gene names and its solution

Background

Since whole genome sequences of rice were made publically accessible, the number of articles on new rice genes has increased remarkably. The Committee on Gene Symbolization, Nomenclature and Linkage (CGSNL) of the Rice Genetics Cooperative published the gene nomenclature system for rice and encouraged researchers to follow the rules before publishing their results. The CGSNL provides an on-line registration system for newly identified rice genes to prevent conflicts and/or duplication of gene name in journal articles.

Findings

Recently, the CGSNL surveyed genes in the rice WRKY family in published journal articles and found several duplicated gene names.

Conclusions

To discuss and resolve inconsistencies in WRKY gene nomenclature, the rice WRKY working group was established and redefined the nomenclature. This report announces the conclusion.     

Characterization of the DNA nucleotide sequences in the genome of red sea bream iridoviruses isolated in Korea

The nucleotide sequences of DNA fragments amplified by polymerase chain reaction (PCR) from four different genomic regions of nine red sea bream iridoviruses (RSIVs) isolated from different species of fish, different areas and in different years in Korea were compared with the reported reference sequences. One isolate, RSIV Namhae, showed 100% homology to the reference sequences, while the other eight isolates, which appeared to contain identical nucleotide sequences, showed 96.6–98.9% homology with reference sequences depending upon the target regions of PCR gene amplification. However, differences in nucleotide sequences were not apparent between the RSIVs isolated in different locations, in different years or in different host species. We also cloned and sequenced the 3′ end flanking region (K1) of the DNA polymerase (DPOL) gene using the cassette ligation-mediated PCR method. This sequence was 4436-bp long and possessed two open reading frames (ORF-1 and ORF-2) oriented in opposite directions. The putative proteins encoded by these two ORFs could not be characterized by comparison with the proteins of other species in the data banks. The presence of the ribonucleotide reductase small subunit (RNRS) gene at the 3′ end of the K1 region allowed us to determine that these two genes, RNRS and DPOL, are separated 5508 bp and oriented in the same direction in the genome of RSIV. Moreover, it is of interest that a PstI-restriction fragment, of which the sequence but not the location within the RSIV genome had previously been reported, is located at nucleotide positions from 1096 to 2054, extending from within the ORF-1 region, spanning the intervening sequence between ORF-1 and ORF-2, and extending into the ORF-2 region. Various repeating sequences up to 86 bp were present at the 3′ ends of ORFs, especially within the nucleotide sequences at the 3′ terminus of ORF-2. No similarities were detected when the DNA sequences of the K1 region were compared to the DNA sequences of a repetitive element in the genome of other iridoviruses.     

水稻内稃扭曲突变体palea distortion 1(pd1)的鉴定与基因定位

在对籼稻缙恢10进行EMS(ethyl methane sulfonate)处理后的群体中,发现一个花器官突变体,主要表现为内稃扭曲并呈现外稃化特征,浆片数目增加且呈现稃状特征,雄蕊数目减少至1～4个,部分雄蕊的花丝呈现浆片化特征,暂将其命名为水稻颖壳扭曲突变体palea distortion 1(pd1)。遗传分析表明该突变性状受一个隐形单基因控制。利用群体分离分析法(bulked segregation analysis,BSA),将PD1基因定位在第2染色体的RM13693和RM13936之间,遗传距离分别为3.25cM和3.90cM。该研究结果为PD1基因的图位克隆奠定了基础。     

陆地棉(G. hirsutum L.) N-乙酰氨基葡萄糖转移酶基因(GhGnT)的克隆及耐盐性功能分析

在耐盐相关基因芯片基础上,我们筛选到盐诱导显著上调表达(Ratio>2)的N-乙酰氨基葡糖转移酶基因.选取耐盐材料中9806为材料,根据耐盐性抑制消减文库EST序列设计引物,利用RACE及RT-PCR技术获得该基因cDNA全长1970 by,命名为GhGnT.通过Blast比对,发现该基因与蓖麻乙酞氨基葡萄糖转移酶基因...     

利用农杆菌介导的共转化法获得转基因水稻

以水稻粳稻品种空育131、垦稻12为材料,用农杆菌共转化的方法将质粒p1300和p13W8导入受体材料。结果显示：两种混合共转化菌液按体积比1∶1时共转化效率最高,植株转化效率达4.08%和2.63%。将转化植株进行PCR鉴定,13株转基因植株均能检测到质粒p1300的信号,4株检测到质粒p13W8的信号,反义蜡质基因的阳性株占转基因植株总数的30.77%。     

中华蜜蜂GnRH相关受体的基因克隆及表达研究

[目的]克隆中华蜜蜂(Apis cerana cerana)促性腺激素释放激素(gonadotrophin releasing hormone,GnRH)相关受体基因,对其进行表达研究。[方法]利用RT-PCR技术克隆了中华蜜蜂的GnRH相关受体基因的cDNA序列,并对其进行生物信息学分析和表达产物的原位杂交组织化学研究。[结果]序列分析显示,GnRH相关受体的cDNA序列全长1 177 bp,开放阅读框长1 050 bp,编码349个氨基酸残基。推导的氨基酸序列具有7个跨膜区;预测的分子量和等电点分别为40.6 kD和9.54。聚类分析显示该蛋白质与其他已知昆虫的GnRHRⅡ具有较近的亲缘关系。原位杂交显示GnRH相关受体基因在中华蜜蜂的脑、精巢、脂肪体以及肠道等组织均有表达。[结论]该研究结果表明GnRH相关受体为昆虫提供了生殖和新陈代谢之间的分子纽带,其在协调昆虫的生殖活动和环境状况的关系中执行了一定的功能。     

差减cDNA文库技术在植物抗性研究中的应用

综述了差减cDNA文库在植物抗性研究中的应用。