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�й����ﻯѧ���������ѧ�� 2003, Vol. 19 Issue (06 ):698-698     DOI:
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Fusion Expression, Purification of Human NDRG1 and Antibody Preparation
ZHANG Jian 1) , LIU Xin ping 1)* , ZHANG Wei 1) , WU Guo qiang 1) , SHEN Lan 1) , WANG Li feng 1) , SU Jin 1) , ZHANG Wan hui 2) , YAO Li bo 1) ( 1) Department of Biochemistry and Molecular Biology, Fourth Military Medical University, Xi��an 710032, China; 2) Department of Physiology , Fourth Military Medical University, Xi��an 710032, China)

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ժҪ NDRG1����N mycȱ�ݵ�С����̥��֯�з��ֵ�һ�쳣�߱����»��� .���о���ͬ�Ͱ��װ���Ѫ֢����������Ӳ���Ļ���ʱ ,����������꾲����Ƥϸ�� (HUVEC)�з�������NDRG1 .Ϊ���о���NDRG1�Ĺ����Լ��ṹ�빦��֮��Ĺ�ϵ ,��RT PCR���� ,��������RNA�п�¡NDRG1cDNA ,�������вⶨ�� ,��NDRG1����pPROEXHTb��������� ,ת��E .coliDH5������̬ϸ�� ,����LB������л�ø�Ч���ܱ�� .���� 6His NDRG1�ںϵ��׾�Ni2 + NTAż������֬���鴿�� ,ͨ��Բ��ɫ�Է���������ṹ ,���Ϊ :������ :2 3 6 ,��Ƭ�� :1 8 6 ,ת�� :2 5 7,�޹���� :32 0 .�ô��ںϱ��ĵ������߼��� ,�Ʊ��õ���Ч�۵Ŀ��� ,���ù̶���������ά��Ĥ�ϵ�NDRG1��ԭ�׺������Ѫ������˿���������� ,Ϊ��һ���о�NDRG1�Ĺ��ܴ��������õĻ�
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�ؼ���� NDRG1   N-myc   �ںϱ��   Ni2+-NTA   Բ��ɫ��   ����Ѫ��   �����Ʊ�     
Abstract�� NDRG1 is a new gene which expresses abnormally highly in N myc mutant mouse embryos. Human NDRG1 is isolated from human umbilical vein endothelial cells in a study of homocysteine promoting arteriosclerosis diseases. In order to investigate the function and structure function relationship, NDRG1 was expressed in E.coli and the fusion protein was obtained. The RT PCR was used to clone human NDRG1 cDNA from the total RNA of brain and then the gene was sequenced. The recombinant vector NDRG1 pP RO EX HTb was transformed into E.coli DH5, and the strains highly expressing soluble 6His NDRG1 in LB medium were obtained. The fusion protein was purified by Ni 2+ NTA agarose beads. The secondary structure of purified 6His NDRG1 fusion protein was analyzed by circular dichroism. The results indicated that 6His NDRG1 was composed of helix:23 6, ��\|sheet:18 6,turn:25 7 and random:32 0. Immunization in rabbits with the fusion protein was used to obtain the high titer antibody against NDRG1. The specific antibody was purified by absorbing antiserum with NDRG1 antigen immobilized NC filter. These works lay a favorable foundation for the study of the NDRG1 structure and function.
Keywords�� NDRG1   N myc   fusion expression   Ni 2+ NTA   circular dichroism   immune sera   antibody preparation      
Received 2002-11-20;
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�Ž�.��NDRG1���ںϱ������������Ʊ� [J]  �й����ﻯѧ���������ѧ��, 2003,V19(06 ): 698-698
.Fusion Expression, Purification of Human NDRG1 and Antibody Preparation [J]  Chinese Journal of Biochemistry and Molecular Biol, 2003,V19(06 ): 698-698
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http://cjbmb.bjmu.edu.cn/CN/     ��     http://cjbmb.bjmu.edu.cn/CN/Y2003/V19/I06 /698