人工染色体是人工构建的含有天然染色体基本功能单位的载体系统总称。人工染色体是非常优良的载体, 具有超大的接受外源片段能力。 由于不用整合到宿主基因组中, 因此不会引起宿主基因的插入失活, 及抑制转基因表达的位置效应。人工染色体已经从最初的酵母人工染色体(Yeast artificial chromosome, YAC)发展到细菌人工染色体(Bacterial artificial chromosome, BAC), 再扩展到人类人工染色体(Human artificial chromosome, HAC)和植物人工染色体(Plant artificial chromosome, PAC)。文章就这4种人工染色体, 尤其是植物人工染色体的研究进展和应用局限进行综述。目前, YAC和BAC已经广泛应用于基因组图谱制作、序列测定和基因克隆; HAC和PAC在基因治疗、外源医用蛋白的生产、新型优质高产高抗转基因作物构建中显现出广阔的应用前景。随着合成生物学的高速发展, 美国科学家报道合成了一个“人造生命”。但是, 和人工染色体一样, 所谓的“人造生命”, 都是应用最新的基因工程技术, 将不同的生命基础元件拼接组装而成, 脱离了细胞环境并不能够自由存在。
Artificial chromosomes (ACs) are genetic-engineered vector systems with defined native chromosomal elements. ACs have large carrying capacity and genetic stability without integration into host genome, thus avoiding random insertion and positional effects. ACs were first successfully developed in yeast (Yeast artificial chromosome, YAC), and then in bacterium (Bacterial artificial chromosome, BAC), human (Human artificial chromosome, HAC), and plant (Plant artificial chromosome, PAC). Here, we summarized recent progress on ACs, especially, on PAC. To date, YAC and BAC have been widely applied in genome sequencing and gene isolation, while HAC and PAC have been subjected to gene therapy, protein production, and plant transgenesis, respectively. Recently, American scientists reported a man-made genome of prokaryote Mycoplasma mycoides. However, like ACs, this man-made genome was also genetic-engineered product and can’t survive as an independent life without a cellular environment.
[1] Mehta GD, Agarwal MP, Ghosh SK. Centromere identity: a challenge to be faced. Mol Genet Genomics, 2010, 284(2): 75-94.
[2] Dalal Y, Bui M. Down the rabbit hole of centromere assembly and dynamics. Curr Opin Cell Biol, 2010, 22(3): 392-402.
[3] Jain D, Cooper JP. Telomeric strategies: means to an end. Annu Rev Genet, 2010, 44: 243-269.
[4] Giraud-Panis MJ, Pisano S, Poulet A, Le Du MH, Gilson E. Structural identity of telomeric complexes. FEBS Lett, 2010, 584(17): 3785-3799.
[5] Murray AW, Szostak JW. Construction of artificial chromosomes in yeast. Nature, 1983, 305(5931): 189-193.
[6] Burke DT, Carle GF, Olson MV. Cloning of large segments of exogenous DNA into yeast by means of artificial chromosome vectors. Science, 1987, 236(4803): 806-812.
[7] Bellis M, Gérard A, Charlieu JP, Marçais B, Brun ME, Viegas-Péquignot E, Carter DA, Roizès G. Construction and characterization of a partial library of yeast artificial chromosomes from human chromosome 21. DNA Cell Biol, 1991, 10(4): 301-310.
[8] Green ED, Riethman HC, Dutchik JE, Olson MV. Detection and characterization of chimeric yeast artificial-chromosome clones. Genomics, 1991, 11(3): 658-669.
[9] Roberts L. Two chromosomes down, 22 to go. Science, 1992, 258(5079): 28-30.
[10] Sakata K, Antonio BA, Mukai Y, Nagasaki H, Sakai Y, Ma-kino K, Sasaki T. INE: a rice genome database with an integrated map view. Nucleic Acids Res, 2000, 28 (1): 97-101.
[11] Shizuya H, Birren B, Kim UJ, Mancino V, Slepak T, Tachiiri Y, Simon M. Cloning and stable maintenance of 300-kilobase-pair fragments of human DNA in Escherichia coli using an F-factor-based vector. Proc Natl Acad Sci USA, 1992, 89(18): 8794-8797.
[12] Shizuya H, Kouros-Mehr H. The development and applications of the bacterial artificial chromosome cloning system. Keio J Med, 2001, 50(1): 26-30.
[13] Kim UJ, Shizuya H, Kang HL, Choi SS, Garrett CL, Smink LJ, Birren BW, Korenberg JR, Dunham I, Simon MI. A bacterial artificial chromosome-based framework contig map of human chromosome 22q. Proc Natl Acad Sci USA, 1996, 93(13): 6297-6301.
[14] Asakawa S, Abe I, Kudoh Y, Kishi N, Wang YM, Kubota R, Kudoh J, Kawasaki K, Minoshima S, Shimizu N. Hu-man BAC library: construction and rapid screening. Gene, 1997, 191(1): 69-79.
[15] Harrington JJ, Van Bokkelen G, Mays RW, Gustashaw K, Willard HF. Formation of de novo centromeres and construction of first-generation human artificial microchromosomes. Nat Genet, 1997, 15(4): 345-355.
[16] Irvine DV, Shaw ML, Choo KH, Saffery R. Engineering chromosomes for delivery of therapeutic genes. Trends Biotechnol, 2005, 23(12): 575-583.
[17] Heller R, Brown KE, Burgtorf C, Brown WR. Mini-chromosomes derived from the human Y chromosome by telomere directed chromosome breakage. Proc Natl Acad Sci USA, 1996, 93(14): 7125-7130.
[18] Duncan A, Hadlaczky G. Chromosomal engineering. Curr Opin Biotechnol, 2007, 18(5): 420-424.
[19] Lufino MM, Edser PA, Wade-Martins R. Advances in high-capacity extrachromosomal vector technology: episomal maintenance, vector delivery, and transgene expression. Mol Ther, 2008, 16(9): 1525-1538.
[20] Macnab S, Whitehouse A. Progress and prospects: human artificial chromosomes. Gene Therapy, 2009, 16(10): 1180-1188.
[21] Keith KC, Copenhaver GP, Luo S, Preuss D. Arabidopsis centromeres: minichromosome analysis of functional centromere domains. Abstract of 11th Arabidopsis Meeting, 2000, 41: 49.
[22] Carlson SR, Rudgers GW, Zieler H, Mach JM, Luo S, Grunden E, Krol C, Copenhaver GP, Preuss D. Meiotic transmission of an in vitro-assembled autonomous maize minichromosome. PLoS Genet, 2007, 3(10): 1965-1974.
[23] Onomura K, Kurata N. Construction of rice
