Glyco-poly-L-lysine is better than liposomal delivery of exogenous genes to rat liver

AIM To compare the effects of liposomes andglyco-poly-L-lysine on liver targeted uptake andexpression of plasmid in rat liver.METHODS After binding with lipofectamine orgalactose-terminal glyco-poly-L-lysine,theplasmid could be expressed in eukaryotic cellswhen injected into Wistar rats by intravenousroute.At different time intervals after the injection,the distribution and expression of the plasmid inliver of rats were observed and compared using insitu hybridization and immunohistochemistry.RESULTS The expression of the plasmid bindingto liposomes or G-PLL could be markedly observed24 h later,and began to decrease one week later,but it still could be observed up to three weeks.Both liposomes and G-PLL could deliver theplasmid to the liver effectively,but the effect of thelatter was better than the former concerning thedistribution and expression of the plasmid targeteduptake in the liver.CONCLUSION G-PLL is better than liposome asthe targeted carrier for delivering exogenous genesto the liver.

Fluorescence Tracking of Exogenous DNA in Genetic Transformation of the Chinese Oak Silkmoth Antheraea Pernyi via Sperm-Mediated Gene Transfer

Exogenous DNA expressing green fluorescent protein( GFP) and labeled with fluorescein isothiocyanate( FITC) was used to transform the Chinese oak silkmoth Antheraea pernyi( A. pernyi)via sperm-mediated gene transfer( SMGT). Sperms entry into the female reproductive system and eggs were observed using fluorescence microscopy. The ability of A. pernyi sperms to uptake exogenous DNA was confirmed,and transfer of the exogenous DNA was shown by GFP expression in the transgenic eggs. Our result suggested that SMGT could also be used to directly generate transgenic A. pernyi expressing functional genes of interest.

Reproductive characteristics of transgenic (TG) chickens caping an exogenous gene

An exogenous gene (lacZ/MiwZ) introduced into the germinal crescent region (GCR) of avian embryos was con-firmed to be successfully transferred to the gonads via the primordial germ cells (PGCs). Following hatching, the chickswere raised until the stage of sexual maturation. The incorporation of MiwZ DNA was detected in male and female trans-genic chickens, respectively. The normal male and female transgenic birds were subjected to artificial insemination ac-cording to routine methods. Fertilized eggs obtained from female transgenic chickens were incubated for 72 h and the em-bryos removed from the yolk were examined by X-gal staining to detect the introduction of MiwZ in the offspring. As aresult, the expression of MiwZ was detected in the offspring. Furthermore, the presence of MiwZ in the extracts fromembryos was also detected by polymerase chain reaction (PCR) analysis. In male transgenic chickens, the presence of in-jected MiwZ in the extracts from sperm was also confirmed. The exogenous gene introduced into the GCR migrated suc-cessfully to the gonad resulting in its incorporation into the offspring and spermatozoa of transgenic chickens. (Asian JAndrol 1999 Sep ; 1: 139 - 144)

Exogenous reference gene normalization for real-time reverse transcription-polymerase chain reaction analysis under dynamic endogenous transcription

Quantitative real-time reverse transcription-polymerase chain reaction （qPCR） is widely used to investigate transcriptional changes following experimental manipulations to the nervous system. Despite the widespread utilization of qPCR, the interpretation of results is marred by the lack of a suitable reference gene due to the dynamic nature of endogenous transcription. To address this inherent deficiency, we investigated the use of an exogenous spike-in mRNA, luciferase, as an internal reference gene for the 2ct normalization method. To induce dynamic transcription, we systemically administered capsaicin, a neurotoxJn selective for C-type sensory neurons expressing the TRPV-1 receptor, to adult male Sprague-Dawley rats. We later isolated nodose ganglia for qPCR analysis with the reference being either exogenous luciferase mRNA or the commonly used endogenous reference 13-111 tubulin. The exogenous luciferase mRNA reference clearly demonstrated the dynamic expression of the endogenous reference. Furthermore, variability of the endogenous reference would lead to misinterpretation of other genes of interest. In conclusion, traditional reference genes are often unstable under physiologically normal situations, and certainly unstable following the damage to the nervous system. The use of exogenous spike-in reference provides a consistent and easily implemented alternative for the analysis of qPCR data.

Effects of Exogenous Ethylene and 1-Methylcyclopropene on of DpXTH Gene Expression in Dahlia(Dahlia pinnata Cav.) Flowers

Objectives] This study was conducted to investigate the regulatory mechanism of ethylene on DpXTH gene expression in dahlia （ Dahlia pinnata Cav.） flower petals.[Methods] Dahlia flower petals were treated with ethephon （ETH） and 1-methylcyclopropene （1-MCP） at different flower development stages and for different durations in vivo . Then, the relative expression of DpXTH1 and DpXTH2 in flower samples in different treatments was quantified with real-time PCR, using β-actin as a reference gene.[Results] The expression of DpXTH1 and DpXTH2 in ETH- or 1-MCP-treated dahlia flower petals increased at first, and decreased subsequently with increasing duration of treatment. The relative expression of both DpXTH1 and DpXTH2 in ETH-treated samples reached the maximum level at 12 h. Moreover, the relative expression of both DpXTH1 and DpXTH2 in dahlia flower petals treated with ETH at early stage of flowering was higher than that of the samples treated at other flowering stages; in 1-MCP-treated samples, it was higher at full-flowering stage than at other stages. The maximum relative expression levels of DpXTH1 and DpXTH2 in ETH- and 1-MCP-treated samples were all higher than those of control.[Conclusions] Exogenous ethylene can accelerate the opening of dahlia flowers, and 1-MCP can inhibit this process, so they can be used to regulate development of dahlia flowers.

The establishment of stable transfection of human breast cancer cell line MDA-MD-468 with exogenous PTEN gene

To investigate exogenous PTEN gene transfected human breast cancer cell line MDA-MD-468.Methods Using the lipofectamine 2000 transfection technique,wild type PTEN gene was transducted into an in vitro cultured highly metastatic breast cancer cell line MDA-MD-468.After transfection,the cells were selected by G418.The resistant clones were chosen and expanded in DMEM culture medium.RT-PCR,immunohistochemical method and western blot were used to determine the expression of target genes.Results An anti-G418 cell clone was established and expanded in culture.The transfected PTEN gene MDA-MD-468 cells showed expression of PTEN mRNA and PTEN protein.Conclusion Human breast cancer cell line MDA-MB-468 established in this study expresses consistently exogenous PTEN genes.4 refs,6 figs.

Biological effect of expression of exogenous human nuclear receptor hLRH-1 in SW480 cells and its molecular mechanism

Objective： To explore the possible biological function of human nuclear receptor hLRH-1 in tumorigenesis and progress of colon cancer. Methods： Plasmids pcDNA3-hLRH-1 were introduced into SW480 cells via lipofectamine. The expression of mRNA and protein of exogenous hLRH-1 were detected by RT-PCR and western blotting, respectively. MTT assay was carried out to survey the proliferation of SW480 cells with overexpression of hLRH-1. Meanwhile, the expression of proliferation-related genes cyclin E1 and cyclin D1, and apoptosis-related genes PTEN and Rbl, were analyzed by realtime RT-PCR. Results： The proliferation of SW480 cells was promoted under the condition of overexpression of hLRH-1. The expression of cyclin E1 was up-regulated significantly, while that of PTEN and Rbl were down-regulated in SW480 cells with overexpressed hLRH-1. Conclusion： The expression of exogenous hLRH-1 in SW480 cells induced the proliferation resulting form up-regulation of cyclin E1, as well as participated in the regulation of apoptosis via influencing the expression of PTEN and Rb1.

Transgenic Crops by Direct Treatment of Exogenous DNA without Agrobacterium tumefaciens Plasmid and Tissue Culture

Gene transfer methods are developing quickly recently, but each method has its limitations. We introduce a new gene transfer technique in this paper, which is simple, effective, and easy to operate,but does not get enough attention from scientists. This technique is used to transform plants by in jecting exogenous DNA to stigma, style, ovary, young fruit or meristem of the recipient, or soaking the recipient's seeds in exogenous DNA solution. Lots of heritable variations were found in many characters of many crops. It may be used to create new germplasms or realize gene exchange between different species, genera, or families, even between animals and plants. A brief discussion was given to the mechanism of exogenous DNA introduction, integration into and expression in the recipient. We also discussed the merits and limitations of the technique.Currently there are two successful approaches that can be used to transform paints genetically,but each method has its limitations that are delaying the application of the techniques to certain commercially important crops. The first technique exploits a natural genetic engineer, Agrobacterium tumefaciens, which contains a tumor-inducing (Ti) plasmid that transfers a DNA segment (the T-DNA) from the plasmid to the nuclear genome of infected plants (or in vitro to plant tissue). The method is restricted to dicotyledenous plants; monocotyledenous plants are usually not susceptible to agrobacterial infection. The second technique involves direct transfer of DNA to plant protoplast, prepared by enzymatic digestion of cell walls, for example by chemically stimulated uptake using polyethylene glycol or a high voltage pulse, generating transient 'holes' in the protoplast membrane. This technique depends on a tissue culture system that allows regeneration of mature plants from protoplasts. But so far it is impossible to achieve plant regeneration from protoplasts in many crops. Both techniques use dominant selectable markers (for example, kanamycin resistance) to select for the transformed tissue or plant which can then be screened for expression of co-transferred but unselected genes (Lichenstein, 1987).Now there is a new successful method which can transform various crops, regardless of dicots or monocots, cereals or legumes. It doesn't need Agrobacterium tumefaciens and plasmid, doesn't depend on the tissue culture system that allows regeneration of mature plants from protoplasts.Comple and advance equipments are not necessary. It is very simple, but very effective. Next is a review about the technique, its application in several crops, the mechanism of transformation, and its merits and limitations.

EXPRESSION OF rhBMP-7 GENE IN TRANSDUCED BONE MARROW DERIVED STROMAL CELLS

OBJECTIVE: To explore the possibility of expression of exogenous gene in transduced bone marrow derived stromal cells (BMSCs). METHODS: The marker gene, pbLacZ, was transferred into cultured BMSCs and the expression of transduced gene by X-gal staining was examined. Then plasmid pcDNA3-rhBMP7 was delivered to cultured BMSCs. Through immunohistochemical staining and RT-PCR assay, the expression of rhBMP7 gene was detected. RESULTS: The exogenous gene could be expressed efficiently in transduced BMSCs. CONCLUSION: The present study provided a theoretical basis to gene therapy on the problems of bone and cartilage tissue.

转录组测序分析外源水杨酸诱导茶树热激蛋白基因的响应

水杨酸是诱导植物抗性机制中重要的信号分子,外源喷施水杨酸能够调控多种防御相关蛋白质,提升农作物的抗病能力。开展外源水杨酸诱导茶树抗性机制的研究能够挖掘抗病基因,为茶树抗病育种提供分子基础。本研究采集外源喷施水杨酸0 h、6 h、12 h、24 h、48 h的茶树叶片进行转录组测序与分析,结果表明,外源喷施水杨酸6 h、12 h、24 h、48 h时茶树叶片内差异表达基因数量分别为9 360个、3 399个、596个、115个,外源水杨酸处理后各时间点均发生差异表达的基因共604个。KEGG功能富集结果显示,处理后6 h时富集于植物激素信号转导、植物-病原菌互作、核糖体、剪接体和碳代谢通路上的差异表达基因数量分别为95个、73个、121个、94个、154个。差异表达基因中有12个热激因子基因、40个热激蛋白基因和12个WRKY家族转录因子基因上调表达。处理48 h后,无上调表达的热激因子基因,但仍有28个热激蛋白基因上调表达。病程相关蛋白基因在检测阶段均上调表达。外源水杨酸的诱导作用在处理6 h时最为明显,并且引起了大量热激蛋白的响应。本研究结果为开展外源水杨酸诱导茶树抗病机制和茶树抗病分子育种研究提供了参考。

外源抗生素抗性基因在农田系统中的定殖机制综述

外源抗生素抗性基因(Antibiotic resistance genes,ARGs)对土壤生态健康和农产品质量安全构成严重威胁,但ARGs在土壤-农作物系统中定殖机理不清、跨界迁移机制不明,直接制约了农业系统中细菌耐药污染的有效防控。本文简要论述农田系统ARGs目前赋存现状,系统综述外源ARGs在农田土壤和农作物中的动态演变特征及其与土壤特性的关联性,梳理了ARGs在土壤中侵入-持留-迁移的定殖分子机制以及揭示耐药菌与内生菌之间的交互过程及驱动因素,并提出关于未来农田ARGs环境行为和阻控研究的展望和建议。

外源蔗糖对甘蓝型油菜种子萌发与幼苗发育的影响

萌发调控是促进甘蓝型油菜快速出苗、齐苗的重要基础,为探索外源蔗糖对甘蓝型油菜种子萌发与幼苗发育的影响,选用早熟油菜品种湘油420进行了不同条件下萌发试验,检测了水分、脱落酸、内源糖及关键基因的表达变化。结果表明:外源蔗糖延缓湘油420种子萌发启动过程,提高萌发整齐度,在子叶展开后促进子叶转绿、真叶形成,抑制主根过度伸长同时促进侧根发育。外源蔗糖对湘油420种子萌发过程中水分吸收表现先抑制后促进,播种后3 h内抑制水分吸收,并在播种后6 h内解除吸水抑制;萌发过程中种子内ABA含量持续降低,外源蔗糖促进播种后3 h内ABA含量的下降;播种后15~18 h内可溶性总糖含量迅速降低,还原性糖含量在萌发初期随萌发进程增加,外源蔗糖对可溶性总糖含量变化无明显影响,但抑制播种后9 h还原糖的增加、促进播种15 h后还原糖含量增加;萌发过程中2个蔗糖磷酸合酶(BnaSPS)基因BnaC09g37470D和BnaA10g15120D相对表达量在胚根萌出阶段显著降低、子叶转绿后表达回升,外源蔗糖抑制胚根萌出前2个BnaSPS基因表达,在子叶转绿后显著诱导其中BnaC09g37470D基因表达。综上,研究初步揭示了外源蔗糖通过渗透性作用调节吸涨起始阶段水分吸收,通过糖代谢调节萌发阶段种子内源糖利用和幼苗发育初期糖类转化而延缓甘蓝型油菜种子萌发启动、促进吸涨后种子萌发进程而提高种子萌发整齐度、促进幼苗发育的多重作用。

谷子CPP基因家族鉴定及外源硒响应分析

【目的】探究谷子CPP基因家族成员特征及在外源硒处理下的响应模式,为谷子富硒高叶酸新品种选育提供遗传材料。【方法】借助生物信息学工具鉴定谷子CPP家族成员,用qRT-PCR技术对CPP基因在不同组织及外源硒处理下的表达水平进行检测。【结果】(1)谷子基因组中包含9个CPP基因,定位于6条染色体上,分别命名为SiCPP1-SiCPP9,所有成员均定位在细胞核,蛋白质二级结构中无规则卷曲占比最重。(2)谷子CPP蛋白可分为4个亚家族,相同亚家族之间保守基序和结构域的数目及分布相似。(3)启动子分析发现谷子CPP家族中存在大量光、生长发育、激素及胁迫响应元件。(4)谷子CPP家族成员在根、茎、叶和穗中差异表达,SiCPP5、SiCPP6、SiCPP7和SiCPP8对外源硒响应最强烈。【结论】谷子CPP家族成员具有组织表达特异性,且对外源硒存在不同程度响应。

中华鳖卵黄蛋白原基因克隆与表达及外源E2的影响

为探究中华鳖(Pelodiscus sinensis)卵黄蛋白原Vtg基因的生物学信息与表达及其作为生理与毒理研究标志物的可行性,本实验克隆获得VtgAB1、VtgAB2、VtgAB3(GenBank登录号OK287927、OK287929、OK287928)基因序列,其cDNA长分别为5055、5537、5306 bp,分别编码了1684、1760、1725个氨基酸。三种亚型Vtg蛋白均由脂蛋白N-末端(Vitellogenin_N)、1943DUF、1944DUF、PV、D型血管血友病因子(VWD)5个保守结构域构成,为完整型Vtg。同源性分析发现皆与龟鳖目具有更近的亲缘关系,同时在3种亚型中VtgAB2与VtgAB3更相似。荧光定量PCR显示,3种亚型Vtg只在雌性肝脏中强烈表达。在60日龄、2龄、4龄雌性个体肝脏中,Vtg表达量随年龄呈上升趋势,VtgAB1、VtgAB2为主要表达亚型。原位杂交结果显示,3种亚型Vtg在卵原细胞内未发现杂交信号;在生长期卵母细胞胞质和滤泡细胞中发现杂交信号;成熟期卵母细胞胞质中未发现杂交信号,在滤泡细胞和周围的网状组织中发现杂交信号。信号在初级卵母细胞的细胞质中分布较为均匀且表达最强,随着卵母细胞的发育信号聚集在核周区且信号逐渐减弱。以雄性中华鳖为对象,注射雌激素E2,3种亚型Vtg表达量表现出明显的时间和剂量相关性。研究结果表明,在中华鳖卵黄积累过程中Vtg发挥重要作用,首次证实其Vtg存在内源性合成,并发现其可以作为雌激素暴露的生物标志物。

禽传染性支气管炎病毒感染性克隆的快速构建及应用

为构建高效且易于操作传染性支气管炎病毒(IBV)基因组的反向遗传学平台,本研究将IBV H120株全基因组经RT-PCR分段扩增,并在其基因组的5'端引入人巨细胞病毒(CMV)启动子序列以及3'端引入丁型肝炎病毒核酶序列(HDVRz)和牛生长激素(BGH)多聚腺苷酸化信号序列(后续称HB序列),获得H120株基因组的9个片段(CMV-F1、F2、F3、F4、F5、F6、F7、F8和F9-HB);其中片段F7通过引物引入沉默突变C20356T作为遗传标记。采用转化偶联重组(TAR)克隆技术将9个DNA片段与线性质粒p YES1L转化酿酒酵母细胞,快速构建H120株基因组全长cDNA克隆,经连接PCR(Junction-PCR)筛选及测序鉴定获得感染性克隆质粒p YES1L-H120。为了提高病毒拯救效率,同时构建表达IBV N蛋白的辅助质粒pcDNA3.1-N+3'UTR。将p YES1L-H120与pcDNA3.1-N+3'UTR共转染BHK-21细胞,48 h后收获细胞,接种9日龄SPF鸡胚盲传3代,经胚体病变观察和测序显示获得IBV反向遗传重组病毒r H120株。为了以IBV为载体表达外源基因,本研究将病毒基因组的5a基因替换为eGFP基因,采用TAR克隆技术构建以IBV为载体表达外源e GFP基因的cDNA克隆pYES1L-H120-Δ5a-eGFP,与辅助质粒共转染BHK-21细胞,随后接种SPF鸡胚拯救嵌合病毒r H120-Δ5a-eGFP,并采用PCR和测序鉴定该重组病毒正确构建。结果显示,采用TAR克隆技术能够在短时间内获得IBV反向遗传重组病毒且快速操作病毒基因组获得以IBV为载体稳定表达外源基因的嵌合病毒。本研究首次建立了基于TAR克隆技术的IBV反向遗传操作平台,为研究病毒的生物学特性、载体疫苗的开发和抗病毒药物的筛选提供高效便捷的工具。

茄子PLC基因家族鉴定及其对低磷胁迫的响应研究

磷脂酶C(PhospholipaseC,PLC)是磷脂代谢中的主要水解酶之一,在植物生长发育和胁迫响应的信号转导过程中起重要作用。本研究从茄子基因组中鉴定出9个PLC基因,分布在6条染色体上,可分为两个亚家族,分别命名为SmPI-PLC1~SmPI-PLC6和SmNPC1~SmNPC3,同亚族的PLC成员在基因结构和保守结构域上表现出一定的相似性;SmPLC启动子中存在多个响应逆境、光形态建成和激素信号的顺式作用元件;茄子的4条染色体与拟南芥3条染色体存在共线性,5条染色体与番茄4条染色体存在共线性。茄子组织特性分析表明,SmNPC2在其种子、根和叶中的表达量均高于茎、花和果实;与处理0d相比,随低磷胁迫时间延长,茄子幼苗根中SmNPC2表达量呈先升高后降低趋势,处理后1d,其表达量升高3.22倍,差异显著;添加外源生长素后,其表达量与处理0d时无显著差异。综上,本研究从全基因组水平鉴定和分析了茄子PLC基因家族,为深入研究其功能及解析低磷胁迫下的作用机制提供了参考依据。

Molecular mechanism of mureidomycin biosynthesis activated by introduction of an exogenous regulatory gene ssa A into Streptomyces roseosporus

Mureidomycins(MRDs), a group of unique uridyl-peptide antibiotics, exhibit antibacterial activity against the highly refractory pathogen Pseudomonas aeruginosa. Our previous study showed that the cryptic MRD biosynthetic gene cluster(BGC) mrd in Streptomyces roseosporus NRRL 15998 could not be activated by its endogenous regulator 02995 but activated by an exogenous activator Ssa A from sansanmycin’s BGC ssa of Streptomyces sp. strain SS. Here we report the molecular mechanism for this inexplicable regulation. EMSAs and footprinting experiments revealed that Ssa A could directly bind to a 14-nt palindrome sequence of 5′-CTGRCNNNNGTCAG-3′ within six promoter regions of mrd. Disruption of three representative target genes(SSGG-02981, SSGG-02987 and SSGG-02994) showed that the target genes directly controlled by Ssa Awere essential for MRD production. The regulatory function was further investigated by replacing six regions of SSGG-02995 with those of ssa A.Surprisingly, only the replacement of 343–450 nt fragment encoding the 115–150 amino acids(AA) of Ssa A could activate MRD biosynthesis. Further bioinformatics analysis showed that the 115–150 AA situated between two conserved domains of Ssa A.Our findings significantly demonstrate that constitutive expression of a homologous exogenous regulatory gene is an effective strategy to awaken cryptic biosynthetic pathways in Streptomyces.

重组火鸡疱疹病毒活载体疫苗研究进展

火鸡疱疹病毒因具有基因组庞大、复制非必需区域多、容纳外源基因能力强、安全性好、免疫持续时间长、相关基因操作技术成熟等众多优点成为活载体疫苗研究的热点。利用同源重组、细菌人工染色体、Fosmid和CRISPR等技术,研究人员成功构建了火鸡疱疹病毒的感染性克隆,并将其转染至易感细胞中拯救重组病毒。为使重组病毒高效表达外源蛋白,通常选择合适的插入位点、启动子和终止序列,并对外源基因序列进行优化处理。目前,以火鸡疱疹病毒为载体的重组疫苗在禽流感、新城疫、传染性法氏囊病、传染性喉气管炎、鹦鹉热衣原体病、艾美尔球虫病和鸭霍乱等多种疾病的预防和控制中展现出了广阔的应用前景。本文从火鸡疱疹病毒活载体疫苗的发展历程、构建方法与应用实践等方面进行综述,旨在为相关领域研究提供理论和实践指导。

外源性胰岛素自身免疫综合征合并线粒体糖尿病1例并文献复习

目的报道1例罕见的外源性胰岛素自身免疫综合征(exogenous insulin autoimmune syndrome,EIAS)合并线粒体糖尿病(mitochondrial diabetes mellitus,MDM)患者诊断、治疗的过程。方法回顾南部战区总医院2018年12月收治的1例以EIAS合并MDM的发病过程、病史及相关辅助检查,总结分析其明确诊断、治疗的过程及特殊性。结果本例患者30岁发病,糖尿病史7年余,初期即出现耳鸣、听力下降,既往于外院曾应用多种胰岛素剂型,在治疗过程中曾出现低血糖,注射部位皮肤出现水泡,伴皮肤瘙痒,同时出现胰岛素水平显著升高,且与C肽结果“分离”,胰岛素自身抗体(insulin autoantibody,IAA)阳性,考虑诊断为EIAS。治疗上停用胰岛素药物,予二甲双胍、DPP-Ⅳ抑制剂、阿卡波糖等口服降糖药物,胰岛素逐渐降至正常,血糖控制尚可;但患者糖尿病治疗过程中逐渐出现神经系统症状,头磁共振提示脑干和双侧小脑半球萎缩,查阅相关文献及结合患者病史,完善基因检测,最终明确患者为线粒体DNA MT-TL1基因A3243G突变所致的MDM。同时检测患者乳酸偏高,二甲双胍可能加重乳酸堆积,甚至诱发乳酸酸中毒,故停用二甲双胍降糖治疗,同时加强饮食和运动控制,患者定期规律门诊随访,血糖控制尚可。结论本例患者初诊糖尿病后应用外源性胰岛素制剂诱发EIAS,但因疾病不断进展,出现无法解释的神经系统症状,完善基因检测明确为MDM。

微小杆菌冷激蛋白(CSP)基因密码子偏好性分析

对微小杆菌(Exobacterium profundaum)冷激蛋白(CSP)基因(EpCSP)密码子偏好性进行分析,为后续EpCSP基因的功能验证及遗传转化等研究提供理论依据。结果表明:EpCSP基因的密码子偏好性较弱,偏好使用A/T结尾的密码子;比较分析不同物种CSP基因的密码子偏好性参数发现,大多数物种CSP基因密码子偏好使用G/C结尾的密码子;21个物种CSP基因的CDS序列及RSCU值聚类分析表明,微小杆菌与另外20个不同物种CSP基因密码子使用偏好性存在一定差异;中性分析、PR2-plot分析、ENc-plot等都证实了自然选择是影响CSP基因密码子偏好性形成的最主要原因;与7种模式生物的基因组密码子使用频率对比研究表明,在筛选异源转化受体时,作为真核表达系统的酿酒酵母比作为原核表达系统的大肠杆菌更利于EpCSP基因异源表达,而在选择植物遗传转化受体时,拟南芥和小麦比烟草和水稻更适合作为EpCSP基因的遗传转化受体。本研究初步探索了EpCSP基因密码子使用偏好性,对以后开展基因功能验证有重要的指导作用,同时也可为研究微小杆菌分子进化提供一定的理论基础。