Methods we used the foreskin tissue as a cell sourse , harvested and cultivated the melanocytes and keratinocytes respectively , and then constructed the mixed cultivating model of them 方法以包皮组织作为细胞来源，分别培养角质形成细胞、黑素细胞；体外构建黑素细胞与角质形成细胞直接接触的混合培养模型。
Dh5 a . methods : the target gene of the domain was obtained by rt - pcr from cultured human keratinocytes coming from isolation and culture of human skin . the target gene fragment with 647 bp was loaded into cloning vector pgem - t and transforming e . coli 方法：通过对人类皮肤组织的分离与培养，利用rt - pcr技术从培养角质形成细胞获取该功能区目的基因，并将长647bp的目的基因片段装入克隆载体pgem - t中，转化大肠杆菌dh5 ，经酶切鉴定及测序证实为目的片段后，再将目的片段装入表达载体pbv - 220转化大肠杆菌dh5进行原核表达。
Ii ) two fragments ( about 240bp and 130bp ) were amplified from human keratinocytes total rna by rt - pcr . the recombinant pm - hpabl and pm - hpabs plasmids were constructed by inserting 240bp and 130bp pcr products into pmd 18 - t vector , respectively . the recombinants were identified by restriction enzyme analysis and dna sequencing , iii ) two orfs ( > 100bp ) were found in the insert sequence of pm - hpabl 应用smartpcr试剂盒和简并引物从人皮肤角质形成细胞cdna中扩增到长分别为24obp和13obp左右的2种片段，将它们插入pmd18一t载体，用酶切法初步筛选阳性重组子pm . hrabl和pm一hrabs ，对阳性重组子进一步作测序鉴定。
