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Mar Biotechnol (NY). 2019 Apr;21(2):206-216. doi: 10.1007/s10126-018-09873-8. Epub 2019 Jan 10.

Construction of a Genomic Bacterial Artificial Chromosome (BAC) Library for the Prawn Macrobrachium rosenbergii and Initial Analysis of ZW Chromosome-Derived BAC Inserts.

Author information

1
Key Laboratory of Freshwater Aquatic Genetic Resources, Ministry of Agriculture, National Demonstration Center for Experimental Fisheries Science Education, Shanghai Engineering Research Center of Aquaculture, Shanghai Ocean University, 201306, Shanghai, People's Republic of China.
2
College of Life Science and Technology, Huazhong Agricultural University, 430070, Wuhan, People's Republic of China.
3
Key Laboratory of Freshwater Aquatic Genetic Resources, Ministry of Agriculture, National Demonstration Center for Experimental Fisheries Science Education, Shanghai Engineering Research Center of Aquaculture, Shanghai Ocean University, 201306, Shanghai, People's Republic of China. gfqiu@shou.edu.cn.
4
College of Fisheries and Life Science, Shanghai Ocean University, 999 Hucheng Huan Road, Pudong New Area, 201306, Shanghai, People's Republic of China. gfqiu@shou.edu.cn.

Abstract

Knowledge on sex determination has proven valuable for commercial production of the prawn Macrobrachium rosenbergii due to sex dimorphism of the male and female individuals. Previous studies indicated that prawn sex is determined by a ZW-ZZ chromosomal system, but no genomic information is available for the sex chromosome. Herein, we constructed a genomic bacterial artificial chromosome (BAC) library and identified the ZW-derived BAC clones for initial analysis of the sex chromosomal DNA sequence. The arrayed BAC library contains 200,448 clones with average insert size of 115.4 kb, corresponding to ∼ 4× coverage of the estimated 5.38 Gb genome. Based on a short female-specific marker, a Z- and a W-fragment were retrieved with the genomic walking method. Screening the BAC library using a ZW-specific marker as probe resulted in 12 positive clones. From these, a Z-derived (P331M17) and a W-derived (P122G2) BAC clones were randomly selected and sequenced by PacBio method. We report the construction of a large insert, deep-coverage, and high-quality BAC library for M. rosenbergii that provides a useful resource for positional cloning of target genes, genomic organization, and comparative genomics analysis. Our study not only confirmed the ZW/ZZ system but also discovered sex-linked genes on ZW chromosomes for the first time, contributing to a comprehensive understanding of the genomic structure of sex chromosomes in M. rosenbergii.

KEYWORDS:

BAC; Genome size; Macrobrachium rosenbergii; Sex chromosome

PMID:
30632018
DOI:
10.1007/s10126-018-09873-8
[Indexed for MEDLINE]

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