优质蛋白玉米胚乳贮存蛋白积累的电泳分析

玉米胚乳的２２ ｋＤ和２０ ｋＤ醇溶蛋白在授粉后１５ 天开始积累,编码２２ ｋＤ 和２０ ｋＤ醇溶蛋白的结构基因在胚乳发育过程中同时表达。优质蛋白玉米和ｏ２ 玉米的胚乳中,２２ ｋＤ和２０ ｋＤ醇溶蛋白的合成受到抑制,即ｏ２ 基因对２２ ｋＤ和２０ ｋＤ醇溶蛋白的合成有负的调节作用。Ｍｏ１７／ｏ２ 和Ｍｏ１７ 胚乳醇溶蛋白的双向电泳结果表明,Ｍｏ１７／ｏ２ 的２７ ｋＤ、２２ ｋＤ、２０ ＫＤ和１５ ｋＤ醇溶蛋白的合成均受到强烈的抑制。遗系０４１／ｏ２ 和遗系０４０／ｏ２ 胚乳醇溶蛋白的双向电泳结果表明,二者只在高分子量的蛋白质斑点区域有一些细微的差别。可溶性蛋白的ＳＤＳ－ＰＡＧＥ分析表明,Ｍｏ１７／ｏ２ 胚乳的可溶性蛋白比其同型系Ｍｏ１７ 少３８．７ ｋＤ 和２６．７ ｋＤ两条谱带,多２７．２ ｋＤ和２６．１ ｋＤ两条谱带。二者出现的可溶性蛋白的差异是ｏ２ 基因调控的结果。遗系０４１／ｏ２ 胚乳的可溶性蛋白比其同型系０４０／ｏ２ 多１８．６ ｋＤ和１７．６ ｋＤ两条谱带,少４０．２ ｋＤ 一条谱带,这与ｏ２ 基因修饰因子的作用密切关联     

Electromicrographic Analysis Storage Protein Accumulation in Endosperms of Maize with Qualified Protein

The characteristics of storage protein accumulation of maize with qualified protein (MQP) and 02 maize were analysed basing on the genetical and biochemical point of views. The 22 kD and 20 kD zeins in the developing endosperms of maize accumulated 15 days after pollination. The structural genes encoding 22 kD and 20 kD zeins in the developing endosperms were simutaneously expressed. In the endosperms of MPQ and o2 maize the synthesis of 22 kD and 20 kD zeins was suppressed. That is to say, o2 gene negatively regulated the synthesis of 22 kD and 20 kD zeins. Two-dimentional electrophoretic analysis of zeins in the maize endosperms further revealed the effects of o2 gene and its modifiers on the synthesis of zeins. In Mol7 and Mo17/o2 endosperms the synthesis of 27 kD, 22 kD, 20 kD and 15 kD zeins was severely suppressed. In 041/oz and 040/o2 endosperms little difference existed SDS-PAGE analysis of the soluble proteins of Mol 7 and Mo17/o2 endosperms revealted that two bands with molecular weight (MW) of 38.7 kD and 26.7 kD were present in wild type but absent in o2 mutant, while two bands with MW 27.2 kD and 26.1 kD were present in o2 mutant but absent in wild type. These differences were resulted from the effect of o2 gene. In 040/02 and 041/o2 endosperms two bands with MW 18.6 kD and 17.6 kD were present in 041/o2 but absent in 040/02 while one band with MW 40. 2 kD was present in 040/02 and absent in 041/o2, which was closely related to the effects of the modifiers of o2 gene.     

N端融合外源蛋白的兔出血症病毒衣壳蛋白自聚能力的研究

将兔出血症病毒衣壳蛋白VP6 0基因插入杆状病毒转移载体pBLUEBACHIS2_B的 6 HIS表达标签下游 ,与线性化野生型杆状病毒基因组DNA共转染Sf9昆虫细胞 ,经蚀斑纯化后获克隆化重组杆状病毒pBLUEBACHIS2B_VP6 0。以重组杆状病毒感染Sf9细胞 ,经SDS_PAGE和Westernblot检测显示高效表达一分子量为 6 9kD的重组蛋白 ,并且该蛋白可被兔抗RHDV高免血清识别。血凝试验表明 ,该重组蛋白可以凝集人“O”型红细胞 ,血凝价达 2 1 6 ,同时 ,该血凝性可被抗RHDV的高免血清所抑制。经电镜观察 ,重组病毒表达的融合有 6 HIS表达标签的衣壳蛋白仍可在昆虫细胞内自聚成不包裹核酸的、与天然RHDV病毒粒子在物理形态上相似的病毒样颗粒 (VLPs) ,并且该VLPs与兔抗RHDV高免血清作用后于电镜下可见凝集成团的现象 ,表明其与天然RHDV病毒粒子在抗原性上也极为相似     

家蚕溶茧酶基因的真核表达及其产物的生物活性检测

为了研究家蚕 Bombyx mori 溶茧酶基因 （cocoonae）真核表达及其产物的生物活性,将溶茧酶基因（GenBank 登录号 EF428980）克隆至杆状病毒转移载体 pFastBac^TM 1 中获得 pFast-cocoonase,将其转化 DH10Bac 感受态细胞后,PCR 方法检测证实所分离的重组病毒 DNA 中含有目的片段溶茧酶基因。用脂质体法 转染家蚕 BmN 细胞,获得重组病毒。SDS-PAGE 分析显示,感染重组杆状病毒 Bac-cocoonase 的细胞表达产物在约为 27.6 kD 处出现特异性条带,这与预测的蛋白大小相符。用该表达产物与茧丝反应后,电镜下观察茧丝的形态,结果表明表达产物对茧丝的丝胶层有一定的水解作用。     

菜心和水稻绿叶中不同等电点的乙醇酸氧化酶

The proteins with glycolate oxidase activity from B.parachinensis Bailey and rice( Oryza sativa )green leaves were prepared respectively.From the second protein peak on DEAE\|Cellulose column,two glycolate oxidases,expressed as B.parachinensis Bailey GO Ⅲ(specific activity natove 13 2 U·mg -1 ·min -1 )and rice GOⅢ(specific activity 8 8 U·mg -1 ·min -1 ),could not migrate anywhere in 4%～20% native\|PAGE under a pH8.3 buffer system.GOⅢ's p I was about pH8.3. The protein containing B.parachinensis Bailey GOⅢ showed 67±2,43±2,and 38±2 kD in SDS PAGE,band 43±2 kD was the subunit of B.parachinensis Bailey GOⅢ.From the two proteins above,another group of glycolate oxidases,expressed as B.parachinensis Beiley GOⅠ(specific activity 5 U·mg -1 ·min -1 )and rice GOⅠ(specific activity 1 2 U·mg -1 ·min -1 ),showed only one 43±2 kD band in SDS\|PAGE,and was purified on the Sepharose\|6B column which migrated towards anode in the same native\|PAGE showing the M r about 420 kD,or 460 kD and 260 kD respectively.GOⅠ's p I was smaller than pH8.3.Antibody against B.parachinensis Bailey GOⅠ was prepared and its efficacy was about 1/1600 in ELISA.By native\|PAGE,Western blot and rocket immunoelectrophoresis,the third group of glycolate oxidases,expressed as B.parachinensis Bailey GOⅡ and rice G0Ⅱ,were confirmed in crude protein of green leaves and migrated towards cathode under the same native\|PAGE,so GOⅡ's p I was higher than pH8.3.The M r of B.parachinensis Bailey GOⅡ was about 669 kD determined by native\|PAGE Western blot.Rice GOⅠ,rice GOⅢ and rice GOⅡ showed different quantitation under different physiological conditions.Rice GOⅡ could be induced by glycolate.     

Nitric Oxide Inducing Function and Intracellular Movement of Chicken Interleukin-18 in Cultured Cells

To evaluate the characteristics of chicken interleukin-18 (ChIL-18) in different forms in vitro,the ChIL-18 full-length gene (ChIL-18-F) and the ChIL-18 presumed mature protein gene (ChIL-18-M) werecloned and inserted into the eukaryotic expression vector pCI,to construct recombinant pCI-ChIL-18-Fand pCI-ChIL-18-M.The recombinant plasmids were then transferred into chicken splenic lymphocytes(CSLs).Western blot showed that ChIL-18-F,with a molecular weight of 23.0 kDa,was produced in CSLstransfected by pCI-ChlL-18-F;ChIL-18-M,with a molecular weight of 19.5 kDa,was produced in CSLstransfected by pCI-ChIL-18-M.The nitric oxide (NO) level in the transfected CSLs and the culture mediumat different time points was further examined under confocal microscopy using 4.5-diaminofluoresceinstaining.The results showed that both pCI-ChIL-18-F and pCI-ChIL-18-M groups showed significantincrease in intracellular and extracellular NO production compared with pCI transfected control cells.Theseresults suggest that both ChIL-18-F and ChIL-18-M could stimulate NO secretion in CSLs.To characterizethe intracellular distribution of ChIL-18,ChIL-18-F and ChIL-18-M were each fused to the enhanced greenfluorescent protein gene,and expressed in Vero cells.The results showed that the ChIL-18-F tended to themembranous region in Vero cells,while ChIL-18-M did not.This indicates that the N-terminal 27 amino acidpeptide helped ChIL-18 target to Veto cell membranes.     

基于猪细小病毒病毒样颗粒的结肠癌靶向纳米载体的构建

【目的】获得具有结肠靶向的纳米载体。【方法】采用SOE-PCR方法将具有结肠靶向的TK肽序列插入到猪细小病毒(PPV)结构蛋白VP2的环2和环4区域得到TK-vp2(?vp2)基因,在Bac-to-Bac?杆状病毒表达系统中构建、表达和自组装。【结果】通过SOE-PCR方法扩增获得?vp2基因,在Bac-to-Bac?杆状病毒表达系统中构建得到Bacmid-?vp2,经脂质体转染至Sf9昆虫细胞得到重组杆状病毒。直接免疫荧光试验、SDS-PAGE和Western blot检测结果表明?VP2蛋白在Bac-to-Bac?杆状病毒表达系统中获得融合表达,目的蛋白约70 k D;透射电子显微镜结果显示?VP2能自组装形成病毒样颗粒(TK-VLPs),直径范围在22 nm-30 nm。【结论】获得纳米载体TK-VLPs,为进一步研究其作为结肠靶向的纳米载体奠定物质基础。     

Nitric oxide inducing function and intracellular movement of chicken interleukin-18 in cultured cells

To evaluate the characteristics of chicken interleukin-18 （ChIL-18） in different forms in vitro, the ChIL-18 full-length gene （ChIL-18-F） and the ChIL-18 presumed mature protein gene （ChIL-18-M） were cloned and inserted into the eukaryotic expression vector pCI, to construct recombinant pCI-ChIL-18-F and pCI-ChIL-18-M. The recombinant plasmids were then transferred into chicken splenic lymphocytes （CSLs）. Western blot showed that ChIL-18-F, with a molecular weight of 23.0 kDa, was produced in CSLs transfected by pCI-ChIL-18-F; ChIL-18-M, with a molecular weight of 19.5 kDa, was produced in CSLs transfected by pCI-ChIL-18-M. The nitric oxide （NO） level in the transfected CSLs and the culture medium at different time points was further examined under confocal microscopy using 4,5-diaminofluorescein staining. The results showed that both pCI-ChIL-18-F and pCI-ChIL-18-M groups showed significant increase in intracellular and extracellular NO production compared with pCI transfected control cells. These results suggest that both ChIL- 18-F and ChIL- 18-M could stimulate NO secretion in CSLs. To characterize the intracellular distribution of ChIL-18, ChIL-18-F and ChIL-18-M were each fused to the enhanced green fluorescent protein gene, and expressed in Vero cells. The results showed that the ChIL-18-F tended to the membranous region in Veto cells, while ChIL- 18-M did not. This indicates that the N-terminal 27 amino acid peptide helped ChIL-18 target to Vero cell membranes.     

Purification and characterization of a thermostable beta-galactosidase from kidney beans (Phaseolus vulgaris L.) cv. PDR14

Using five different steps, beta-Galactosidase has been purified from kidney beans to apparent electrophoretic homogeneity with approximately 90-fold purification with a specific activity of 281 units mg-1 protein. A single band was observed in native PAGE. Activity staining of the native gel with 5-bromo 4-chloro 3-indoxyl beta-D-galactopyranoside (X-Gal) at pH 4.0 also produced a single band. Analytical gel filtration in Superdex G-75 revealed the molecular mass of the native protein to be approximately 75 kD. 10% SDS-PAGE under reducing conditions showed two subunits of molecular masses, 45 and 30 kD, respectively. Hence, beta-galactosidase from kidney beans is a heterodimer. A typical protein profile with lambda max at 280 nm was observed and A280/A260 ratio was 1.52. The N-terminal sequence of the 45 kD band showed 86% sequence homology with an Arabidopsis thaliana and 85% with Lycopersicon esculentum putative beta-galactosidase sequences. The Electrospray Mass Spectrometric analysis of this band also revealed a peptide fragment that had 90% sequence homology with an Arabidopsis thaliana putative beta-galactosidase sequence. The N-terminal sequencing of the 30 kD band as well as mass spectrometric analysis both by MALDI-TOF and ES MS revealed certain sequences that matched with phytohemagglutinin of kidney beans. The optimum pH of the enzyme was 4.0 and it hydrolysed o- and p-nitrophenyl beta-D galactopyranoside with a Km value of 0.63 mmol/L and 0.74 mmol/L, respectively. The energy of activation calculated from the Arrhenius equation was 14.8 kcal/mol enzyme site. The enzyme was found to be comparatively thermostable showing maximum activity at 67 degrees C. Thermal denaturation of the enzyme at 65 degrees C obeys single exponential decay with first order-rate constant 0.105 min-1. Galactose, a hydrolytic product of this enzyme was a competitive inhibitor with a Ki of 2.7 mmol/L.     

Highly efficient and economical baculovirus expression system for preparing human papillomavirus type16 virus-like particle

To improve the existing human papillomavirus type16 (HPV16) virus-like particle (VLP) preparation, a highly efficient, economical and timesaving system was established. Sf-9 cells were infected with recombinant baculovirus containing the target gene encoding HPV16L1 protein with 6xHis tag, and harvested 72 h postinfection (p.i.) at 27 degrees. The ProBond(TM) purification system was used for protein purification. The molecular weight of expressed HPV16L1 protein was 58 kD as revealed by SDS-PAGE, and confirmed by Western blot. The purity of denatured and native HPVL1 proteins that were prepared were 91.9% and 71.5%, respectively, which corresponded to a yield of 2.26 mg denatured protein and 1.84 mg native protein per 2x10(7) cells. The proteins were further analyzed by mouse erythrocyte hemagglutination assay and hemagglutination inhibition assay, and there effects on VLP formation were also visualized by transmission electron microscopy. Results showed that the native protein purified was biologically active as natural HPVL1 protein, inducing the murine erythrocyte agglutination and VLP formation. In addition, the purified recombinant HPV16L1 native protein with 6xHis tag could self-assemble into virions in vitro. Hopefully, the present expression and purification system is promising to be convenient, timesaving and economical for preparation of HPV16 VLP vaccine.     

菠菜乙醇酸氧化酶同工酶的亚基组成

我们首次报告 ,菠菜有ＧＯⅠ (ｐＩ≈ 7.4 )、ＧＯⅡ(ｐＩ≈ 9.4 )和ＧＯⅢ (ｐＩ≈ 8.3 ) 3种乙醇酸氧化酶(ＧＯ)同工酶 ;ＧＯⅠ只含 4 0± 2ｋＤ一种亚基 ,而ＧＯⅡ和ＧＯⅢ的亚基组成未被研究 ;3种同工酶之间均有免疫同源性[1-4 ] .水稻也存在 3种ＧＯ同工酶 ,其中ＧＯⅡ (ｐＩ >8.3 )能被乙醇酸所诱导 用柱层析法纯化可获得经ＳＤＳ ＰＡＧＥ后为 4 3± 2ｋＤ单带的水稻ＧＯⅠ[5～ 7] .以上初步解释了前人报告ＧＯ电荷不均一的原因[5] .最近从菠菜分离得到含ＧＯⅡ的蛋白和含ＧＯⅢ的蛋白 ,其ＳＤＳ ＰＡＧＥ分别为 67± 2ｋＤ和 4 0±…     

急性早幼粒细胞白血病维甲酸耐药性的双向电泳分析

目的:研究急性早幼粒细胞白血病(APL)对全反式维甲酸(ATRA)治疗敏感和耐药患者的外周血淋巴细胞在蛋白质组水平上的差异。方法:采用双向凝胶电泳(2-DE)对敏感和耐药患者的外周血淋巴细胞进行蛋白质组差异分析。结果:ATRA敏感和耐药患者外周血淋巴细胞的2-DE平均蛋白质点分别为(746±57)和(617±41),敏感与耐药患者的2-DE相比,有16个蛋白点表达明显上调,22个明显下调。另有4个蛋白点(Mr/pI:24.6kD/8.05,32.3kD/5.17,22.3kD/6.51,25.1kD/7.09)在敏感患者中特异表达,5个蛋白点(Mr/pI:21.9kD/5.45,23.4kD/6.27,22.9kD/6.65,23.9kD/7.39,24.7kD/7.65)在耐药患者中特异表达。结论:结果提示这些差异表达的蛋白质可能与APL对ATRA耐药的机制有关,该研究有助于揭示APL对ATRA耐药机理和发现新的临床分子标志物。     

HCV核心蛋白在昆虫细胞中的表达及其免疫学特性的初步评价

通过逆转录－聚合酶链反应（ＲＴ－ＰＣＲ）从丙肝患者的血清中分离出编码完整ＨＣＶ核心蛋白（Ｃ区）的ｃＤＮＡ片段,并将其克隆到杆状病毒转移质粒中。重组转移质粒ＤＮＡ与线性的杆状病毒ＤＮＡ共转染Ｓｆ９昆虫细胞,经蚀斑筛选获得了带编码全部核心蛋白基因的重组杆状病毒。重组病毒感染细胞后表达ＨＣＶ核心蛋白,其分子量的为２０ｋＤ。免疫印染和酶联免疫实验表明,此重组蛋白能被人ＨＣＶ阳性血清所识别。动物实验表明此重组蛋白能诱导小鼠产生特异性抗体。     

J亚群禽白血病病毒跨膜蛋白gp37基因克隆与表达

禽白血病病毒(ALV)跨膜蛋白(TM)在病毒-细胞融合过程中起关键作用,其蛋白内部存在的许多高度保守序列有可能成为抗病毒的重要靶点。对TM结构和功能的深入研究将为抗禽白血病病毒乃至其它反转录病毒相关制剂的研制提供科学的理论基础。本研究通过PCR从本实验室分离的ALV-J山东汶上毒株获得表达TM的gp37基因,克隆连接pMD18-T载体并测序,从已构建的质粒pMD18-T-gp37中酶切回收gp37基因,构建重组转移载体pFastBacHTb-gp37。利用昆虫杆状病毒表达系统对gp37基因进行表达,通过间接免疫荧光和Western blot检测gp37基因表达产物,间接免疫荧光显示,重组杆状病毒感染的sf9细胞呈现明显的强阳性反应;Western blot分析,重组病毒感染的sf9细胞蛋白显示出约21kD的阳性条带。结果表明,gp37基因在sf9细胞内表达成功。     

鸡IL-2/NDV-F蛋白抗原表位融合基因的表达及免疫性初探

研究鸡白细胞介素-2(chicken interleukin-2,ChIL-2)的免疫佐剂功能,构建ChIL-2与新城疫病毒F蛋白多抗原表位(NDV-F)的嵌合基因,以对鸡新城疫疫病进行防治。采用重叠延伸PCR方法通过基因柔性接头将ChIL-2基因和NDV-F多抗原表位基因构建成ChIL-2-linker-NDV-F嵌合基因并克隆入PET-32a载体,经测序鉴定后,转化BL21大肠杆菌,IPTG诱导表达6×His融合蛋白,Ni2~+亲和柱纯化,表达产物经SDS-PAGE、Western blot和间接ELISA检测和鉴定。结果表明,实验成功构建并克隆了ChIL-2-linker-NDV-F嵌合基因,嵌合基因在大肠杆菌中表达的ChIL-2-linker-NDV-F融合蛋白分子量约为48kD,表达量约占菌体蛋白总量的45%,纯化后的ChIL-2-linker-NDV-F融合蛋白能与感染NDV的鸡血清发生反应。上述结果证明ChIL-2-linker-NDV-F嵌合基因在原核细胞中能有效表达,且融合蛋白具有较强的特异性和免疫原性。     

猪圆环病毒2型ORF2基因在昆虫细胞中的表达及其特性

利用Bac-to-Bac杆状病毒表达系统将圆环病毒2型的ORF2全基因克隆到杆状病毒转移载体pFastBac^TM1中,获得重组转移载体pFast-OFR2,再将其转化进含穿梭载体Bacmid的感受态细胞DH10Bac中,发生转座作用,经蓝白菌落筛选得到含ORF2基因的重组穿梭载体Bac-ORF2,以脂质体介导的方法将重组穿梭载体转染sf9细胞,获得重组病毒,命名为Ac-ORF2。间接免疫荧光分析表明,PCV2阳性血清能使Ac-ORF2感染的sf9昆虫细胞呈强的荧光着色; SDS-PAGE与Western-blotting分析可见大小约为28kD的特异性带,表明Ac.ORF2在sf9细胞中成功表达了PCV2-ORF2蛋白。将该表达蛋白纯化并经磷钨酸负染后,通过电镜观察可见形态与PCV2病毒粒子相似的病毒样颗粒(VLPs),其中某些颗粒由于中心浓染而似空衣壳,其直径也为17nm左右。     

禽白血病病毒J亚群囊膜蛋白env基因的克隆和表达

禽白血病病毒J亚群（ALV－J）是90年代鉴定出的ALV的新亚群,其囊膜蛋白env基因序列别与ALV A－E亚群的有相当大的差别。为ALV－J env基因及春表达产物的特点,用PCR方法扩增出ADOL－4817毒株的env基因,并克隆进TA载体,经电泳鉴定大小为1.7kb。将克隆出的env基因与杆状病毒pBlue-Bac4表达质粒DNA连接,构建成转移性载体pBac4817env,通过与Bac-N-Blue杆状病毒DNA共转染,区得了重组病毒rBac4817env-2。该重组杆状病毒感染Sf9细胞,能高效表达env基因产物,免疫荧光分析结果证明,单克隆抗体G2或多价兔抗env gp37血清能识别Sf9细胞,能高效表达env基因表达的特异性抗原;Western blotting分析结果表明,表达的重组基因产物的分子量大小约为90kD-94kD。用这些重组基因产物免疫鸡可以诱导鸡导鸡产生出高滴度的抗ALV－J特异性抗体。这一结果提示,这种杆状病毒表达的重组基因产物有助于ALV－J env基因生物学特性的深入研究。     

Baculovirus-mediated expression of the Na+/glucose cotransporter in Sf9 cells.

We have used baculovirus (AcNPV) to express the Na+/glucose cotransporter protein in cultured Sf9 cells. We constructed a baculovirus transfer vector containing the cDNA for the rabbit intestinal Na+/glucose cotransporter (SGLT1) under the control of the polyhedrin gene promoter. Recombinant baculovirus was obtained by cotransfection of SF9 cells with wild-type AcNPV DNA and the transfer vector. Recombinant virus was identified by Southern blotting and then purified. Recombinant infected Sf9 cells expressed a protein which was recognized by anti-peptide antibodies raised to sequences of the cloned Na+/glucose cotransporter. This protein migrated with a molecular mass of 55 kD by SDS-PAGE, similar to the in vitro translation product of SGLT1. An identical protein was metabolically labeled with [35S]methionine. Cells which synthesized the transport protein showed Na(+)-dependent alpha MeGlc transport. Micromolar phlorizin inhibited transport. Uninfected and wild-type virus infected Sf9 cells did not have Na(+)-dependent glucose transport. All transport protein migrated at 45% sucrose (w/w) by density gradient sedimentation, suggesting that the expressed transporter is membrane associated. We conclude that we have functionally expressed the rabbit intestinal Na+/glucose cotransporter in Sf9 cells. The transporter is not heavily glycosylated, and this is consistent with previous work showing that glycosylation is not necessary for function. We are poised to purify and characterize this protein from a structure-function perspective.     

中枢神经系统靶向性CuZn—SOD的构建和表达

SOD对中风等由氧自由基毒性引起的神经性紊乱有保护作用,但因血脑屏障使血液中的SOD不能进入中枢神经系统。靶向性SOD可能是进入该系统的途径之一。将人CuZn-SODcDNA与破伤风毒素C部分基因融合,分别整合进pET－22b（＋）及pFastBacHTb载体中,并分别在E.coli及粉纹夜蛾Tn-5B1-4细胞中表达。表达产物分子量为68kD,与理论计算值。蛋白质印迹实验证实,其表达产物能与人CuZn－SOD多克隆抗人本及抗破伤毒素全毒互抗体有免疫反应。在Tn细胞中高效表达,表达产物占可溶性总蛋白质的20％,表达产物有SOD活性,且具有逆行轴突运输的能力。这为靶向性SOD的进一步应用创造了条件。