Expression of human lactoferrin in the mammary glands of transgenic mice using regulatory elements of rat β-casein gene = 흰쥐 베타-카제인 유전자의 발현조절 부위를 이용하여 유선에서 사람 락토페린을 발현하는 형질전환 생쥐의 개발

Abstract

Two human lactoferrin expression vectors (pCChcLf and pCChcLf-1) were constructed using rat /9-casein gene and human lactoferrin cDNA. The recombinant DNAs containing human lactoferrin cDNA were microinjected into the fertilized eggs of hybrid mice (BDF1 : C57BL x DBA) and the DNA-injected eggs were transferred into the oviducts of foster mothers. Genomic DNAs were isolated from the tails of mice born from the microinjected eggs and analyzed by Southern blot analysis. As a result, 5 and 9 transgenic mice with CChcLf and CChcLf-1 gene were produced, respectively. To determine tissue-specificity of transgene expression, Northern blot analysis was performed. Female transgenic mice were killed at day 10 of lactation and total RNAs from various tissues were isolated. Based on Northern blot analysis, it was shown that transgene was mainly expressed in the mammary glands of transgenic mice. In addition, the hu' man lactoferrin in milk was detected by enzyme-linked immunosorbent assay. For this study, milk was obtained from the mammary glands of the transgenic mice at day 10 of lactation. In line #2 of CChcLf and line 社7 of CChcLf-1 transgenic mice, human lactoferrin was secreted into the milk at concentration levels of 340 ng /ml and 60 ng /ml, respectively.