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利用ＣＤ２３ｃＤＮＡ全基因克隆ｐＵＣＤ９７６，分别以限制性内切酶ＥｃｏＲＩ和ＸｂａＩ酶切该重组质粒ＤＮＡ，回收１．０ｋｂ的ＣＤ２３ｃＤＮＡ全基因、ＨｉｎｄⅢ酶切后回收３’端６０７ｂｐ的结合区段基因以及ＥｃｏＲⅠ和ＨｉｎｄⅡ酶切回收５’端４０３ｂｐ的调控区段基因。将各回收片段先以Ｋｌｅｎｏｗ酶补平后，插入已补平的ｐＺＩＰ逆转录病毒载体ＢａｍＨⅠ单切点，利用地高辛素标记的ＣＤ２３ｃＤＮＡ全基因探针，以斑点核酸杂交筛选逆转录病毒重组体，结合用限制性内切酶ＢａｍＨＩ和／或ＢｇｌⅡ酶切鉴定插入方向，最终获得正、反向插入的ＣＤ２３全基因－逆转录病毒重组体各２个，反向插入３’端和５’端部分基因片段的重组体分别为４个和１个。为进行ＣＤ２３反义ＲＮＡ的研究和真核表达ＣＤ２３提供了可靠的物质基础。
The full-length CD23 cDNA was cloned into pUCD976 to digest the recombinant plasmid DNA with restriction endonucleases EcoRI and XbaI respectively. The full-length CD23 cDNA of 1.0kb was recovered. The HindⅢ digested the 3’-end 607bp binding fragment and the EcoRI and Hind Ⅱ restriction endonuclease recovery 403bp 5 ’regulatory segment gene. The recovered fragments were first filled in with Klenow enzyme, inserted into the flanking pZIP retroviral vector BamHI cleavage site, digoxigenin labeled CD23 cDNA whole gene probe to spot nucleic acid hybridization screening retrovirus recombinant , Combined with restriction endonuclease BamHI and / or Bgl Ⅱ digestion to identify the direction of insertion, the final positive and negative insert CD23 whole gene - retrovirus recombinant two reverse insertion 3 ’and 5’ end Partial gene fragments of the recombinant were 4 and 1. CD23 antisense RNA for the study and eukaryotic expression of CD23 provides a reliable material basis.