With the identification of more than a dozen novel Hermansky-Pudlak Syndrome （HPS） proteins in vesicle trafficking in higher eukaryotes, a new class of trafficking pathways has been described. It mainly consists of ...With the identification of more than a dozen novel Hermansky-Pudlak Syndrome （HPS） proteins in vesicle trafficking in higher eukaryotes, a new class of trafficking pathways has been described. It mainly consists of three newly-defined protein com- plexes, BLOC-l, -2, and -3. Compelling evidence indicates that these complexes together with two other well-known complexes, AP3 and HOPS, play important roles in endosomal transport. The interactions between these complexes form a network in protein trafficking via endosomes and cytoskeleton. Each node of this network has intra-complex and extra-complex interactions. These complexes are connected by direct interactions between the subunits from different complexes or by indirect interactions through coupling nodes that interact with two or more subunits from different complexes. The dissection of this network facilitates the understanding of a dynamic but elaborate transport machinery in protein/membrane trafficking. The disruption of this network may lead to abnormal trafficking or defective organellar development as described in patients with Hermansky-Pudlak syndrome.展开更多
目的:利用全外显子测序技术,对Noonan综合征(NS)患儿及父母进行测序,筛选致病基因突变,探讨基因型与临床表型的关系。方法:提取NS患儿及其父母外周血基因组DNA,使用Agilent Sure Select Human All Exome V6试剂盒进行全基因组外显子捕获...目的:利用全外显子测序技术,对Noonan综合征(NS)患儿及父母进行测序,筛选致病基因突变,探讨基因型与临床表型的关系。方法:提取NS患儿及其父母外周血基因组DNA,使用Agilent Sure Select Human All Exome V6试剂盒进行全基因组外显子捕获,利用Illumina Hiseq X Ten测序仪进行双端高通量测序。通过生物信息学分析,利用db SNP数据库、千人基因组、Ex AC数据库分析变异频率,确定突变位点,用Sanger测序法进行验证。结果:患儿,女性,4岁1个月,生长发育落后、特殊面容。超声心动图示肺动脉瓣狭窄、卵圆孔未闭,临床未给予明确诊断。通过全外显子测序,在患儿BRAF基因中找到一个杂合错义突变c.770A>G(p.Q257R),其父母此位点为正常基因型,该突变为患儿新发突变。结合临床表型和基因检测结果,该患儿最终被诊断为NS。结论:本研究明确了NS和突变位点的关系,强调了全外显子测序用于非典型病例明确诊断和准确遗传咨询的实用性。展开更多
An insect excitatory toxin gene from Buthus martensii Karseh （BmKIT） was cloned into the expression vector, pET-28a. BmKIT was expressed as inclusion bodies in Eseherichia coli BL21 （DE3） host cells. The authentic...An insect excitatory toxin gene from Buthus martensii Karseh （BmKIT） was cloned into the expression vector, pET-28a. BmKIT was expressed as inclusion bodies in Eseherichia coli BL21 （DE3） host cells. The authenticity of in vitro expressed protein was confirmed by Western blot. The inclusion body protein band in SDS-PAGE was excised and the protein, BmKIT, was extracted. Polyelonal antibodies to the purified protein were raised in rabbits. The antibody reacted specifically with the expressed BmKIT and was used to quantify its presence in transgenic cotton.展开更多
基金This work was supported in part by the National Science Fund for Distinguished Young Scholars (No. 30525007)National Basic Research Program of China (No. 2006CB504103+1 种基金 No. 2006CB500704)Hi-Tech Research and Development Program of China (No. 2006AA02Z322)
文摘With the identification of more than a dozen novel Hermansky-Pudlak Syndrome （HPS） proteins in vesicle trafficking in higher eukaryotes, a new class of trafficking pathways has been described. It mainly consists of three newly-defined protein com- plexes, BLOC-l, -2, and -3. Compelling evidence indicates that these complexes together with two other well-known complexes, AP3 and HOPS, play important roles in endosomal transport. The interactions between these complexes form a network in protein trafficking via endosomes and cytoskeleton. Each node of this network has intra-complex and extra-complex interactions. These complexes are connected by direct interactions between the subunits from different complexes or by indirect interactions through coupling nodes that interact with two or more subunits from different complexes. The dissection of this network facilitates the understanding of a dynamic but elaborate transport machinery in protein/membrane trafficking. The disruption of this network may lead to abnormal trafficking or defective organellar development as described in patients with Hermansky-Pudlak syndrome.
文摘目的:利用全外显子测序技术,对Noonan综合征(NS)患儿及父母进行测序,筛选致病基因突变,探讨基因型与临床表型的关系。方法:提取NS患儿及其父母外周血基因组DNA,使用Agilent Sure Select Human All Exome V6试剂盒进行全基因组外显子捕获,利用Illumina Hiseq X Ten测序仪进行双端高通量测序。通过生物信息学分析,利用db SNP数据库、千人基因组、Ex AC数据库分析变异频率,确定突变位点,用Sanger测序法进行验证。结果:患儿,女性,4岁1个月,生长发育落后、特殊面容。超声心动图示肺动脉瓣狭窄、卵圆孔未闭,临床未给予明确诊断。通过全外显子测序,在患儿BRAF基因中找到一个杂合错义突变c.770A>G(p.Q257R),其父母此位点为正常基因型,该突变为患儿新发突变。结合临床表型和基因检测结果,该患儿最终被诊断为NS。结论:本研究明确了NS和突变位点的关系,强调了全外显子测序用于非典型病例明确诊断和准确遗传咨询的实用性。
基金the National Natural Science Foundation of China(No30670282 and 30470239)Natural Science Foundation of Shanxi Province(20051065 and 20041033)Scholar Foundation of Shanxi Province
文摘An insect excitatory toxin gene from Buthus martensii Karseh （BmKIT） was cloned into the expression vector, pET-28a. BmKIT was expressed as inclusion bodies in Eseherichia coli BL21 （DE3） host cells. The authenticity of in vitro expressed protein was confirmed by Western blot. The inclusion body protein band in SDS-PAGE was excised and the protein, BmKIT, was extracted. Polyelonal antibodies to the purified protein were raised in rabbits. The antibody reacted specifically with the expressed BmKIT and was used to quantify its presence in transgenic cotton.
