biologia plantarum

Biologia plantarum 50:131-134, 2006 | DOI: 10.1007/s10535-005-0087-5

Production of human lactoferrin in transgenic cell suspension cultures of sweet potato

S. R. Min¹, J. W. Woo¹, W. J. Jeong¹, S. K. Han¹, Y. B. Lee², J. R. Liu^1,*

¹ Laboratory of Plant Cell Biotechnology, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Yuseong-gu, Daejeon, Korea

² Department of Horticulture, Chungnam National University, Yuseong-gu, Korea

Shoot apical meristem-derived calli were transformed with a hLF cDNA in an attempt to produce human lactoferrin (hLF) in transgenic cell suspension cultures of sweet potato [Ipomoea batatas (L.) Lam.]. Calli were bombarded with tungsten particles coated with the binary vector pLSM1 containing a hLF cDNA under the control of the 35S promoter and the neomycin phosphotransferase gene as a selection marker. Calli were then transferred to Murashige and Skoog (MS) medium supplemented with 4.52 µM 2,4-dichlorophenoxyacetic acid (2,4-D) and 100 mg dm^-3 kanamycin. Kanamycin-resistant calli were selected at four-week intervals and subcultured. Cell suspension cultures were established in liquid MS medium with 4.52 µM 2,4-D. Southern and Northern blot analyses confirmed that hLF cDNA was incorporated into the plant genome and was properly expressed in the cells. ELISA analysis showed that transgenic cells produced hLF up to 3.2 µg mg^-1 (total protein).

Keywords: genetic transformation; Ipomoea batatas; particle bombardment

Subjects: genetic diversity, variability; in vitro culture, cell suspension cultures; Ipomoea batatas; lactoferrin; Northern blot analysis; nutrient medium, Murashige and Skoog; particle bombardment; Southern blot analysis; sweet potato; Western blot analysis

Received: February 23, 2004; Accepted: October 11, 2004; Published: March 1, 2006  Show citation