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유준희(Jun Hi Yoo),정구흥(Gu Hung Jung) 한국식물학회 1994 Journal of Plant Biology Vol.37 No.1
DNA uptake in dry embryos of rice by DNA imbibition was detected by monitoring the expression of chimeric vectors. The selective markers of expression vectors used were β-glucuronidase (GUS) and hygromycin phosphotransferase (HPT) genes under the control of CaMV 35S promoter. Frequency of transient expression of the foreign gene was generally 30-50% varying according to the types of vectors and rice cultivars. Dot blot analysis and DNA sequence analysis of inverse polymerase chain reaction products showed that selected rice in hygromycin B (HmB) medium had HPT gene and CaMV35S promoter DNA sequence in genomic DNA of rice. To investigate what ratio of rice having two marker genes simultaneously as rice embryos imbibed the vector DNA having two HPT and GUS gene, transformants selected in HmB medium were subjected to PCR for GUS gene. It was shown that about 90 percentage of surviving ones in HmB medium had GUS gene.
Aspergillus nidulans 형질전환체의 Genomic DNA 분석
김석준,유준희,정구흥 한국유전학회 1993 Genes & Genomics Vol.15 No.4
Vectors with Clostridium thermosulfurogenes α-amylase gene as a foreign gene and E. coli hph gene as a selectable marker gene which represented hygromycin B resistance(HmB^R) were constructed. AlcA promoter and trpC terminator of Aspergillus nidulans were used on these vectors. Sensitivity of Aspergillus nidulans KCTC6034 to HmB was checked. A. nidulans did not grow at the plates with 600㎍/㎖ of HmB. A. nidulans was transformed with these vectors. Twenty two transformants were obtained and screened by polymerase chanin reaction with genomic DNA to confirm the existence of the foreign DNA. Most of transformants had hph gene sequence and at least 26% of transformants had α-amylase gene sequence.