Skip to main content
Log in

Construction and Characterization of Two Bacterial Artificial Chromosome Libraries of Zhikong Scallop, Chlamys farreri Jones et Preston, and Identification of BAC Clones Containing the Genes Involved in Its Innate Immune System

  • Original Article
  • Published:
Marine Biotechnology Aims and scope Submit manuscript


Large-insert bacterial artificial chromosome (BAC) libraries are necessary for advanced genetics and genomics research. To facilitate gene cloning and characterization, genome analysis, and physical mapping of scallop, two BAC libraries were constructed from nuclear DNA of Zhikong scallop, Chlamys farreri Jones et Preston. The libraries were constructed in the BamHI and MboI sites of the vector pECBAC1, respectively. The BamHI library consists of 73,728 clones, and approximately 99% of the clones contain scallop nuclear DNA inserts with an average size of 110 kb, covering 8.0× haploid genome equivalents. Similarly, the MboI library consists of 7680 clones, with an average insert of 145 kb and no insert-empty clones, thus providing a genome coverage of 1.1×. The combined libraries collectively contain a total of 81,408 BAC clones arrayed in 212 384-well microtiter plates, representing 9.1× haploid genome equivalents and having a probability of greater than 99% of discovering at least one positive clone with a single-copy sequence. High-density clone filters prepared from a subset of the two libraries were screened with nine pairs of Overgos designed from the cDNA or DNA sequences of six genes involved in the innate immune system of mollusks. Positive clones were identified for every gene, with an average of 5.3 BAC clones per gene probe. These results suggest that the two scallop BAC libraries provide useful tools for gene cloning, genome physical mapping, and large-scale sequencing in the species.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Subscribe and save

Springer+ Basic
EUR 32.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or Ebook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others


  • Adachi K, Wakamatsu K, Ito S, Miyamoto N, Kokubo T (2005) An oxygen transporter hemocyanin can act on the late pathway of melanin synthesis. Pigment Cell Research 18:214–219

    Article  PubMed  CAS  Google Scholar 

  • Ashida M, Brey PT (1998) Recent advances on the research of the insect prophenoloxidase cascade. In: Brey PT, Hultmark D (eds) Molecular mechanisms of immune responses in insects. Chapman and Hall, London, p 135e72

    Google Scholar 

  • Chang YL, Tao Q, Scheuring C, Ding K, Meksem K, Zhang HB (2001) An integrated map of Arabidopsis thaliana for functional analysis of its genome sequence. Genetics 159:1231–1242

    PubMed  CAS  Google Scholar 

  • Cunningham C, Hikima J, Jenny MJ, Chapman RW, Fang GC, Saski C, Lundqvist ML, Wing RA, Cupit PM, Gross PS, Warr GW, Tomkins JP (2006) New resources for marine genomics: bacterial artificial chromosome libraries for the Eastern and Pacific oysters (Crassostrea virginica and C. gigas). Mar Biotechnol 8:521–533

    Article  PubMed  CAS  Google Scholar 

  • De Tomaso AW, Weissman IL (2003) Construction and characterization of large-insert genomic libraries (BAC and Fosmid) from the ascidian Botryllus schlosseri and initial physical mapping of a histocompatibility locus. Mar Biotechnol 5:103–115

    PubMed  Google Scholar 

  • Feng J, Vick BA, Lee MK, Zhang HB, Jan CC (2006) Construction of BAC and BIBAC libraries from sunflower and identification of linkage group-specific clones by overgo hybridization. Theor Appl Genet 113:23–32

    Article  PubMed  CAS  Google Scholar 

  • Frijters ACJ, Zhang Z, van Damme M, Wang G, Ronald PC, Michelmore RW (1997) Construction of a bacterial artificial chromosome library containing large EcoR I and Hind III genomic fragments of lettuce. Theor Appl Genet 94:390–399

    Article  CAS  Google Scholar 

  • Guo X, Ford SE, Zhang F (1999) Molluscan aquaculture in China. J Shellfish Res 18:19–31

    Google Scholar 

  • Götz P (1986) Mechanisms of encapsulation in dipteran hosts. In: Lackie AM (ed) Immune mechanisms in invertebrate vectors. Oxford University Press, Oxford, p 1e20

    Google Scholar 

  • He L, Du C, Li Y, Scheuring C, Zhang HB (2007) Large-insert bacterial clone libraries and their applications. In: Liu Z (ed) Aquaculture genome technologies. Blackwell, Ames, pp 215–244

    Chapter  Google Scholar 

  • Li L, Xiang JH, Liu X, Zhang Y, Dong B, Zhang XJ (2005) Construction of AFLP-based genetic linkage map for Zhikong scallop, Chlamys farreri Jones et Preston and mapping of sex-linked markers. Aquaculture 245:63–73

    Article  CAS  Google Scholar 

  • Liu X, Chang Y, Xiang JH, Li F, Liu X, Brovkina EP (2003) Studies on hybridization effects of different geographic population of Chlamys farreri. Acta Oceanol Sin 25:255–263

    Google Scholar 

  • MAC (Ministry of Agriculture of China) Bureau of Aquatic Products. 1986–2001. China Fishery Annual Statistics. Beijing, China

  • Medzhitov R, Janeway CA (1997) Innate immunity, impact on the adaptive immune response. Current Opinion Immunol 9:4e9

    Article  Google Scholar 

  • Nagai T, Kawabata S (2000) A link between blood coagulation and prophenol oxidase activation in arthropod host defense. J Biol Chem 275:29264–29267

    Article  PubMed  CAS  Google Scholar 

  • Pan J, Bao Z, Zhao Y, Wan J, Li Z, Hu X (2002) The genetic diversity of different geographic Zhikong scallop (Chlamys farreri) populations. High Technol Lett 12:78–82

    Google Scholar 

  • Ren C, Xu ZY, Sun S, Lee MK, Wu C, Zhang HB (2005) Genomic libraries for physical mapping. In: Meksem K, Kahl G (eds) The handbook of plant genome mapping: genetic and physical mapping. Wiley-VCH Verlag GmbH, Weinheim, pp 173–213

    Chapter  Google Scholar 

  • Sambrook J, Maniatis T, Fritsch EF (1989) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor

    Google Scholar 

  • Shizuya H, Birren B, Kim UJ, Mancino V, Slepak T, Tachiiri Y, Simon M (1992) 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 89:8794–8797

    Article  PubMed  CAS  Google Scholar 

  • Song L, Li J, Li H, Cui Z, Li C, Xu W, Chang Y (2002a) The genetic structure and genetic differentiation of the natural population and the hatchery stock of Chlamys farreri revealed by RAPD analysis. High Technol Lett 7:83–86

    Google Scholar 

  • Song L, Xu W, Li H1, Li C, Guo X, Xiang JH (2002b) Potential genes involved in immune response identified by expressed sequence tag analysis from scallop Chlamys farreri. In: 5th Symposium on Diseases in Asia Aquaculture, p 141

  • Tao Q, Zhang HB (1998) Cloning and stable maintenance of DNA fragments over 300 kb in Escherichia coliwith conventional plasmid-based vectors. Nucleic Acids Res 26:4901–4909

    Article  PubMed  CAS  Google Scholar 

  • Tao Q, Chang YL, Wang J, Chen H, Islam-Faridi MN, Scheuring C, Wang B, Stelly DM, Zhang HB (2001) Bacterial artificial chromosome-based physical map of the rice genome constructed by restriction fingerprint analysis. Genetics 158:1711–1724

    PubMed  CAS  Google Scholar 

  • Wang S, Bao Z, Pan J (2004) AFLP linkage map of an intraspecific cross in C. farreri. Jof Shellfish Res 23:491–499

    Google Scholar 

  • Wang L, Song L, Chang Y, Xu W, Ni D, Guo X (2005) A preliminary genetic map of Zhikong scallop (Chlamys farreri Jones et Preston 1904). Aquaculture Res 36:643–653

    Article  CAS  Google Scholar 

  • Wang S, Xu P, Thorsen J, Zhu B, de Jong PJ, Waldbieser G, Kucukatas H, Liu Z (2007) Characterization of a BAC Library from Channel Catfish Ictalurus punctatus: Indications of high levels of chromosomal reshuffling among teleost genomes. Mar Biotechnol (Online First)

  • Wu C, Sun S, Padmavathi N, Santos FA, Springman R, Ding K, Meksem K, Lightfoot DA, Zhang HB (2004) A BAC and BIBAC-based physical map of the soybean genome. Genome Res 14:319–326

    Article  PubMed  CAS  Google Scholar 

  • Yang H, Zhang F, Guo X (2000) Triploid and tetraploid Zhikong scallop, Chlamys farreri Jones et Preston, produced by inhibiting polar body I. Mar Biotechnol 2:466–475

    PubMed  CAS  Google Scholar 

  • Zhang H-B (2000) Construction and manipulation of large-insert bacterial clone libraries-manual. Texas A&M University, Texas

  • Zhang H-B, Wu C (2001) BAC as tools for genome sequencing. Plant Physiol Biochem 39:1–15

    Article  Google Scholar 

  • Zhang F, Yang H (1999) Analysis of the causes of mass mortality of farming Chlamys farreri in summer in costal areas of Shandong, China. Mar Sci 1:44–47

    Google Scholar 

  • Zhang H-B, Zhao X, Ding X, Paterson AH, Wing RA (1995) Preparation of megabase-size DNA from plant nuclei. Plant J 7:175–184

    Article  CAS  Google Scholar 

  • Zhang, H-B, Woo S, Wing RA (1996) BAC, YAC and Cosmid Library Construction. In: Foster G, Twell D (eds) Plant Gene Isolation: Principles and Practice. Wiley, England, pp 75–99

    Google Scholar 

  • Zhang L, Bao Z, Cheng J, Li H, Huang X, Wang S, Zhang C, Hu J (2007) Fosmid library construction and initial analysis of end sequences in Zhikong Scallop (Chlamys farreri). Mar Biotechnol 9:606–612

    Article  PubMed  CAS  Google Scholar 

Download references


This project is supported by Key Program of National Natural Science Foundation of China (No. 30730071), Major State Basic Research Development Program of China (2006CB101804), and National High-Tech Research and Development Program of China (2007AA09Z430). The authors thank Prof. Linsheng Song, Prof. Bin Liu, Dr. Bing Wang, Dr. Jiquan Zhang, Miss Fan He, and other laboratory members for technical advice and assistance.

Author information

Authors and Affiliations


Corresponding author

Correspondence to Jianhai Xiang.

Additional information

Yang Zhang and Xiaojun Zhang contributed equally to the article.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Zhang, Y., Zhang, X., Scheuring, C.F. et al. Construction and Characterization of Two Bacterial Artificial Chromosome Libraries of Zhikong Scallop, Chlamys farreri Jones et Preston, and Identification of BAC Clones Containing the Genes Involved in Its Innate Immune System. Mar Biotechnol 10, 358–365 (2008).

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: