太空诱变哈密瓜两性花性状连锁标记的RAPD分析

以哈密瓜品种'早皇后'、太空诱变后两性花株突变体及其后代分离群体为试材,采用BSA方法对哈密瓜两性花性状进行了RAPD分析.研究结果表明:在所筛选的490条10-mer随机引物中,只有S1254在两性花株基因池中扩增到750 bp的多态性条带,而在其它基因型的群体中未扩增到此条带.通过对分离群体及其姐妹系进行单株验证,均获得相同的扩增结果,说明S1254750与两性花性状连锁,该标记与两性花性状的遗传距离为4.5 cM.     

小麦高分子量麦谷蛋白亚基等电点的特性分析

运用IEF×SDS-PAGE和NEPEGE×PAGE 2种双向电泳技术对5个普通小麦品种和1个硬粒小麦品种的高分子量麦谷蛋白亚基等电点的特性进行了研究。结果表明,等位亚基之间的等电点比较相近,而非等位亚基之间的等电点存在不同程度的差异,其中普通小麦G lu-B 1位点的亚基等电点最高,属于碱性类型。用NEPHGE×SDS-PAGE电泳能够对G lu-B 1亚基的等电点特性进行比较分析。通过对麦谷蛋白亚基等电点特性及变异规律进行分析,可以为精确判断亚基类型和鉴定遗传新种质等研究工作提供有价值的信息。     

不结球白菜雄蕊心皮化雄性不育系HGMSⅡ特性研究

对不结球白菜雄蕊心皮化雄性不育系HGMS和HGMSⅡ进行花器形态比较、同功酶分析及结实性研究.结果表明:HGMSⅡ较好地保留了HGMS花瓣萼片化、雄蕊心皮化鲜明特征,而且花瓣EST和POD同功酶酶带与萼片酶带基本相同,心皮化器官EST和POD同功酶酶带与雌蕊酶带相同.与完全雄蕊心皮化的HGMS相比,HGMSⅡ雄蕊心皮化突变主要发生在花药部位,导致花药丧失了产生花粉的能力,从而保留了HGMS不育性稳定、彻底的优良特性.与HGMS相比,HGMSⅡ开花习性明显改善,柱头表面花粉明显增多,雌蕊发育基本正常,单株平均籽粒产量达到8.96 g,较HGMS增加了7.17 g,增幅达401%,结实性得到了显著提高.     

B3(ds-scFv)靶向超抗原的制备及活性鉴定

To construct the expression vector of a recombinant toxin composed of a disulfide stable single-chain antibody from mAbB3 and SEA(D227A),the binding ability and cytotoxicity of the purified renatured products against the B3 positive carcinoma cells was examined. The VH and VL fragments of the mAbB3 were ligated by overlap PCR, the PCR product was cloned to the pET22b expression vector, then the SEA fragment was inserted into the B3dsscFv-pET22b expression vector which was digested by the same restriction enzymes. The expression plasmid was identified by restriction endonucleases digestion and transformed into E.coli BL21(DE3) followed by IPTG induction. The inclusion body was purified through SP-Sepharose cation exchange column after denaturing and refolding and the binding and cytotoxic ability of the purified products was examined by cell-ELISA and non-radioactive cell proliferation assay seperately. The expression vector B3dsscFv-SEA-pET was constructed successfully and the expression product exists mainly in the inclusion body, amounting to 33% of the total protein. The refolding product remains the binding ability of the single-chain antibody and has cytotoxic effect on HT-29 colon carcinoma cells. The stability assay showed that the resulting protein was stable at 37℃. This genetically engineered B3dsscFv-SEA fusion protein has bifunction of tumor targeting and tumor cell killing and promises to be an effective reagent for tumor targeted immunotherapy.